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2023 Competition
IIT-Madras
Commits
f86abf30
Commit
f86abf30
authored
1 year ago
by
Sishir Sivakumar
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Merge branch 'garv' into 'main'
Update notebook.html See merge request
!42
parents
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d0176902
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2 merge requests
!44
Main
,
!42
Update notebook.html
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#342526
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1 year ago
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Stage: deploy
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wiki/pages/notebook.html
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f86abf30
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@@ -5,17 +5,168 @@
{% block page_content %}
<div
class=
"LabNotebook"
<div
style=
"width: 1440px; height: 2782px; position: relative; background: white"
>
<div
style=
"width: 1440px; height: 76px; left: 0px; top: 24px; position: absolute; background: black"
></div>
<div
style=
"width: 1440px; height: 111px; left: 0px; top: 181px; position: absolute; background: #34304D"
></div>
<div
style=
"width: 681px; height: 80px; left: 380px; top: 196px; position: absolute; text-align: center; color: white; font-size: 60px; font-family: Montserrat; font-weight: 700; word-wrap: break-word"
>
Lab Notebook
</div>
<div
style=
"width: 1440px; height: 97px; left: 0px; top: 328px; position: absolute"
>
<div
style=
"width: 1440px; height: 97px; left: 0px; top: 0px; position: absolute; background: #9C93C9"
></div>
<div
style=
"width: 1079px; left: 181px; top: 23px; position: absolute; text-align: center; color: black; font-size: 42px; font-family: Montserrat; font-weight: 700; word-wrap: break-word"
>
Dry Lab
</div>
</div>
<div
style=
"width: 1201px; height: 803px; left: 128px; top: 495px; position: absolute"
>
<div
style=
"width: 200px; height: 785px; left: 0px; top: 0px; position: absolute"
>
<div
style=
"width: 200px; height: 156px; left: 0px; top: 0px; position: absolute"
><span
style=
"color: #221023; font-size: 36px; font-family: Montserrat; font-weight: 700; word-wrap: break-word"
>
June
<br
/></span><span
style=
"color: #221023; font-size: 24px; font-family: Montserrat; font-weight: 400; word-wrap: break-word"
><br
/></span><span
style=
"color: #221023; font-size: 24px; font-family: Montserrat; font-weight: 700; word-wrap: break-word"
>
Week
1
</span></div>
<div
style=
"width: 200px; height: 156px; left: 0px; top: 248px; position: absolute; color: #221023; font-size: 24px; font-family: Montserrat; font-weight: 700; word-wrap: break-word"
>
Week 2
</div>
<div
style=
"width: 200px; height: 156px; left: 0px; top: 424px; position: absolute; color: #221023; font-size: 24px; font-family: Montserrat; font-weight: 700; word-wrap: break-word"
>
Week 3
</div>
<div
style=
"width: 200px; height: 156px; left: 0px; top: 629px; position: absolute; color: #221023; font-size: 24px; font-family: Montserrat; font-weight: 700; word-wrap: break-word"
>
Week 4
</div>
</div>
<div
style=
"width: 1079px; left: 122px; top: 72px; position: absolute"
><span
style=
"color: black; font-size: 24px; font-family: Montserrat; font-weight: 400; word-wrap: break-word"
>
Recruitment
of the team completed
<br
/>
Review of the original work by Salis et al. for better technical understanding of
the problem statement
<br
/>
Review of other work on predictive design of ribosome binding sites - thermodynamic
models, kinetic models, stochastic models, machine learning models
<br
/></span><span
style=
"color: black; font-size: 24px; font-family: Montserrat; font-weight: 400; word-wrap: break-word"
><br
/></span><span
style=
"color: black; font-size: 24px; font-family: Montserrat; font-weight: 400; word-wrap: break-word"
>
Review
of other work on predictive design of ribosome binding sites - thermodynamic models, kinetic models,
stochastic models, machine learning models
<br
/>
Installing the ViennaRNA library for RNA sequence analysis
using Python
<br
/>
Downloading a dataset of 1014 RBS sequences with experimentally determined expression levels
from Reis and Salis (2020)
<br
/></span><span
style=
"color: black; font-size: 24px; font-family: Montserrat; font-weight: 400; word-wrap: break-word"
><br
/></span><span
style=
"color: black; font-size: 24px; font-family: Montserrat; font-weight: 400; word-wrap: break-word"
>
Defining
the interaction between the rRNA and mRNA so as to have minimum hybridisation energy
<br
/>
Including the effect
of spacing between the SD sequence and the start codon
<br
/>
Defining the interaction between the rRNA and the
ribosomal S1 protein
<br
/>
Accounting for the transient regional folding of the mRNA that can hinder RBS
accessibility
<br
/></span><span
style=
"color: black; font-size: 24px; font-family: Montserrat; font-weight: 400; word-wrap: break-word"
><br
/></span><span
style=
"color: black; font-size: 24px; font-family: Montserrat; font-weight: 400; word-wrap: break-word"
>
Including
the presence of standby sites upstream of the SD sequence to which the ribosome can bind if the SD sequence is
transiently folded
<br
/>
Accounting for the effect of various start codons on translation efficiency by using
data from Hecht et al. (2017)
<br
/>
Training and testing different machine learning models to see which works
best for the data at hand
</span></div>
</div>
<div
style=
"width: 1201px; height: 484px; left: 128px; top: 1386px; position: absolute"
>
<div
style=
"width: 200px; height: 478px; left: 0px; top: 0px; position: absolute"
>
<div
style=
"width: 200px; height: 156px; left: 0px; top: 0px; position: absolute"
><span
style=
"color: #221023; font-size: 36px; font-family: Montserrat; font-weight: 700; word-wrap: break-word"
>
July
<br
/></span><span
style=
"color: #221023; font-size: 24px; font-family: Montserrat; font-weight: 400; word-wrap: break-word"
><br
/></span><span
style=
"color: #221023; font-size: 24px; font-family: Montserrat; font-weight: 700; word-wrap: break-word"
>
Week
1
</span></div>
<div
style=
"width: 200px; height: 156px; left: 0px; top: 219px; position: absolute; color: #221023; font-size: 24px; font-family: Montserrat; font-weight: 700; word-wrap: break-word"
>
Week 2
</div>
<div
style=
"width: 200px; height: 156px; left: 0px; top: 279px; position: absolute; color: #221023; font-size: 24px; font-family: Montserrat; font-weight: 700; word-wrap: break-word"
>
Week 3
</div>
<div
style=
"width: 200px; height: 23px; left: 0px; top: 455px; position: absolute; color: #221023; font-size: 24px; font-family: Montserrat; font-weight: 700; word-wrap: break-word"
>
Week 4
</div>
</div>
<div
style=
"width: 1079px; left: 122px; top: 72px; position: absolute"
><span
style=
"color: black; font-size: 24px; font-family: Montserrat; font-weight: 400; word-wrap: break-word"
>
Downloading
a dataset of 16779 RBS sequences characterised by FlowSeq from Reis and Salis (2020) and testing the model on
it
<br
/>
Reading up on different optimisation algorithms that can be used to "mutate" an RBS sequence in silico
in order to achieve a desired expression level
<br
/></span><span
style=
"color: black; font-size: 24px; font-family: Montserrat; font-weight: 400; word-wrap: break-word"
><br
/></span><span
style=
"color: black; font-size: 24px; font-family: Montserrat; font-weight: 400; word-wrap: break-word"
>
Simulated
annealing and gradient descent optimisers were created and tested
<br
/></span><span
style=
"color: black; font-size: 24px; font-family: Montserrat; font-weight: 400; word-wrap: break-word"
><br
/></span><span
style=
"color: black; font-size: 24px; font-family: Montserrat; font-weight: 400; word-wrap: break-word"
>
Optimiser
performance was tested by varying its intrinsic parameters, work is ongoing
<br
/>
A partially working genetic
algorithm was created, work is yet to be finished on this
<br
/>
The two working optimisers were used to
generate RBS sequences with varying expression levels. These sequences will be used in the wet lab using
GFP-expressing Lactococcus lactis to see if the optimisation was successful.
<br
/></span><span
style=
"color: black; font-size: 24px; font-family: Montserrat; font-weight: 400; word-wrap: break-word"
><br
/></span><span
style=
"color: black; font-size: 24px; font-family: Montserrat; font-weight: 400; word-wrap: break-word"
>
All
India iGEM Meet
</span></div>
</div>
<div
style=
"width: 1201px; height: 397px; left: 128px; top: 1958px; position: absolute"
>
<div
style=
"width: 200px; height: 391px; left: 0px; top: 0px; position: absolute"
>
<div
style=
"width: 200px; height: 156px; left: 0px; top: 0px; position: absolute"
><span
style=
"color: #221023; font-size: 36px; font-family: Montserrat; font-weight: 700; word-wrap: break-word"
>
August
<br
/></span><span
style=
"color: #221023; font-size: 24px; font-family: Montserrat; font-weight: 400; word-wrap: break-word"
><br
/></span><span
style=
"color: #221023; font-size: 24px; font-family: Montserrat; font-weight: 700; word-wrap: break-word"
>
Week
1
</span></div>
<div
style=
"width: 200px; height: 156px; left: 0px; top: 132px; position: absolute; color: #221023; font-size: 24px; font-family: Montserrat; font-weight: 700; word-wrap: break-word"
>
Week 2
</div>
<div
style=
"width: 200px; height: 112px; left: 0px; top: 279px; position: absolute; color: #221023; font-size: 24px; font-family: Montserrat; font-weight: 700; word-wrap: break-word"
>
Week 3
</div>
<div
style=
"width: 200px; height: 23px; left: 0px; top: 339px; position: absolute; color: #221023; font-size: 24px; font-family: Montserrat; font-weight: 700; word-wrap: break-word"
>
Week 4
</div>
</div>
<div
style=
"width: 1079px; left: 122px; top: 72px; position: absolute"
><span
style=
"color: black; font-size: 24px; font-family: Montserrat; font-weight: 400; word-wrap: break-word"
>
Genetic
algorithm completed. Found to have the best performance.
<br
/></span><span
style=
"color: black; font-size: 24px; font-family: Montserrat; font-weight: 400; word-wrap: break-word"
><br
/></span><span
style=
"color: black; font-size: 24px; font-family: Montserrat; font-weight: 400; word-wrap: break-word"
>
Attempted
to combine different aspects to the optimization algorithms.
<br
/>
Started work on adding new parameters to the
model to improve accuracy of predictions. Literature review on RNA folding kinetics and RNA degradation was
initiated.
<br
/></span><span
style=
"color: black; font-size: 24px; font-family: Montserrat; font-weight: 400; word-wrap: break-word"
><br
/></span><span
style=
"color: black; font-size: 24px; font-family: Montserrat; font-weight: 400; word-wrap: break-word"
>
Literature
review continued
<br
/></span><span
style=
"color: black; font-size: 24px; font-family: Montserrat; font-weight: 400; word-wrap: break-word"
><br
/></span><span
style=
"color: black; font-size: 24px; font-family: Montserrat; font-weight: 400; word-wrap: break-word"
>
Starting
working with the Kinfold library of ViennaRNA to find the folding time. There was a need to integrate command
line operations with Python code.
</span></div>
</div>
<div
style=
"width: 1201px; height: 339px; padding-bottom: 33.82px; left: 128px; top: 2443px; position: absolute; justify-content: center; align-items: center; display: inline-flex"
>
<div
style=
"width: 1201px; height: 305.18px; position: relative"
>
<div
style=
"width: 226px; height: 305.18px; left: 0px; top: 0px; position: absolute"
>
<div
style=
"width: 226px; height: 146.09px; left: 0px; top: 0px; position: absolute"
><span
style=
"color: #221023; font-size: 36px; font-family: Montserrat; font-weight: 700; word-wrap: break-word"
>
September
<br
/></span><span
style=
"color: #221023; font-size: 24px; font-family: Montserrat; font-weight: 400; word-wrap: break-word"
><br
/></span><span
style=
"color: #221023; font-size: 24px; font-family: Montserrat; font-weight: 700; word-wrap: break-word"
>
Week
1
</span></div>
<div
style=
"width: 200px; height: 146.09px; left: 0px; top: 159.09px; position: absolute; color: #221023; font-size: 24px; font-family: Montserrat; font-weight: 700; word-wrap: break-word"
>
Week 2
</div>
</div>
<div
style=
"width: 1079px; height: 137.66px; left: 122px; top: 67.43px; position: absolute"
><span
style=
"color: black; font-size: 24px; font-family: Montserrat; font-weight: 400; word-wrap: break-word"
>
Kinfold
used to generate folding time data for the dataset.
<br
/>
Prototype for degradation model
built
<br
/></span><span
style=
"color: black; font-size: 24px; font-family: Montserrat; font-weight: 400; word-wrap: break-word"
><br
/></span><span
style=
"color: black; font-size: 24px; font-family: Montserrat; font-weight: 400; word-wrap: break-word"
>
Folding
kinetics and degradation did not improve model performance.
<br
/>
Model finalised without using these
parameters.
</span></div>
</div>
</div>
</div>
<div
class=
"main"
></div>
<div
style=
"
width: 1440px;
height:
4121
px;
height:
5858
px;
position: relative;
background: white;
"
>
<div
class=
"Rectangle7"
style=
"
width: 1440px;
height: 76px;
...
...
@@ -26,7 +177,6 @@
"
></div>
<div
class=
"Rectangle7"
style=
"
width: 1440px;
height: 111px;
...
...
@@ -37,25 +187,25 @@
"
></div>
<div
class=
"LabNotebook"
style=
"
width: 681px;
height: 80px;
left: 3
8
0px;
left: 3
0
0px;
top: 196px;
position: absolute;
text-align: center;
text-align: center;
color: white;
font-size: 60px;
font-family: Montserrat;
font-weight: 700;
word-wrap: break-word;
display: inline;
text-justify: center;
"
>
Lab Notebook
</div>
<div
class=
"FeatureEngineering"
style=
"
width: 1440px;
height: 97px;
...
...
@@ -65,7 +215,6 @@
"
>
<div
class=
"Rectangle11"
style=
"
width: 1440px;
height: 97px;
...
...
@@ -76,7 +225,6 @@
"
></div>
<div
class=
"DryLab"
style=
"
width: 1079px;
left: 181px;
...
...
@@ -90,125 +238,92 @@
word-wrap: break-word;
"
>
Dry
Lab
Wet
Lab
</div>
</div>
<div
class=
"Group42"
style=
"
width: 1
201
px;
height:
803
px;
width: 1
119
px;
height:
2459
px;
left: 128px;
top: 495px;
position: absolute;
"
>
<div
class=
"Group41"
style=
"
width: 200px;
height:
785
px;
height:
156
px;
left: 0px;
top: 0px;
position: absolute;
color: #221023;
font-size: 36px;
font-family: Montserrat;
font-weight: 700;
word-wrap: break-word;
"
>
<div
class=
"JuneWeek1"
August
<br
/>
</div>
<div
style=
"width: 1079px; left: 40px; top: 1183px; position: absolute"
>
<span
style=
"
width: 200px
;
height: 156
px;
left: 0px
;
top: 0px
;
position: absolute
;
color: black
;
font-size: 24
px;
font-family: Montserrat
;
font-weight: 400
;
word-wrap: break-word
;
"
>
<span
style=
"
color: #221023;
font-size: 36px;
font-family: Montserrat;
font-weight: 700;
word-wrap: break-word;
"
>
June
<br
/></span
><span
style=
"
color: #221023;
font-size: 24px;
font-family: Montserrat;
font-weight: 400;
word-wrap: break-word;
"
><br
/></span
><span
style=
"
color: #221023;
font-size: 24px;
font-family: Montserrat;
font-weight: 700;
word-wrap: break-word;
"
>
Week 1
</span
>
</div>
<div
class=
"Week2"
><br
/><br
/></span
><span
style=
"
width: 200px;
height: 156px;
left: 0px;
top: 248px;
position: absolute;
color: #221023;
color: black;
font-size: 24px;
font-family: Montserrat;
font-weight: 700;
word-wrap: break-word;
"
>
Week 2
</div>
<div
class=
"Week3"
>
19 Aug
<br
/></span
><span
style=
"
width: 200px;
height: 156px;
left: 0px;
top: 424px;
position: absolute;
color: #221023;
color: black;
font-size: 24px;
font-family: Montserrat;
font-weight: 700;
word-wrap: break-word;
"
>
Colony PCR (Attempt 1)
</span
><span
style=
"
color: black;
font-size: 24px;
font-family: Montserrat;
font-weight: 400;
word-wrap: break-word;
"
>
Week 3
</div>
<div
class=
"Week4"
: Template solution was made by picking colonies and dispersing in
aq medium.
<br
/></span
><span
style=
"
width: 200px;
height: 156px;
left: 0px;
top: 629px;
position: absolute;
color: #221023;
color: black;
font-size: 24px;
font-family: Montserrat;
font-weight: 700;
word-wrap: break-word;
"
>
Week 4
</div>
</div>
<div
class=
"RecruitmentOfTheTeamCompletedReviewOfTheOriginalWorkBySalisEtAlForBetterTechnicalUnderstandingOfTheProblemStatementReviewOfOtherWorkOnPredictiveDesignOfRibosomeBindingSitesThermodynamicModelsKineticModelsStochasticModelsMachineLearningModelsReviewOfOtherWorkOnPredictiveDesignOfRibosomeBindingSitesThermodynamicModelsKineticModelsStochasticModelsMachineLearningModelsInstallingTheViennarnaLibraryForRnaSequenceAnalysisUsingPythonDownloadingADatasetOf1014RbsSequencesWithExperimentallyDeterminedExpressionLevelsFromReisAndSalis2020DefiningTheInteractionBetweenTheRrnaAndMrnaSoAsToHaveMinimumHybridisationEnergyIncludingTheEffectOfSpacingBetweenTheSdSequenceAndTheStartCodonDefiningTheInteractionBetweenTheRrnaAndTheRibosomalS1ProteinAccountingForTheTransientRegionalFoldingOfTheMrnaThatCanHinderRbsAccessibilityIncludingThePresenceOfStandbySitesUpstreamOfTheSdSequenceToWhichTheRibosomeCanBindIfTheSdSequenceIsTransientlyFoldedAccountingForTheEffectOfVariousStartCodonsOnTranslationEfficiencyByUsingDataFromHechtEtAl2017TrainingAndTestingDifferentMachineLearningModelsToSeeWhichWorksBestForTheDataAtHand"
style=
"width: 1079px; left: 122px; top: 72px; position: absolute"
>
<span
><br
/>
21 Aug
<br
/></span
><span
style=
"
color: black;
font-size: 24px;
font-family: Montserrat;
font-weight: 700;
word-wrap: break-word;
"
>
Gel Check
</span
><span
style=
"
color: black;
font-size: 24px;
...
...
@@ -217,20 +332,25 @@
word-wrap: break-word;
"
>
Recruitment of the team completed
<br
/>
Review of the original work
by Salis et al. for better technical understanding of the problem
statement
<br
/>
Review of other work on predictive design of ribosome
binding sites - thermodynamic models, kinetic models, stochastic
models, machine learning models
<br
/></span
: All PCR products ran on agarose gel; 6, 8, 13, 16 amplified
<br
/></span
><span
style=
"
color: black;
font-size: 24px;
font-family: Montserrat;
font-weight:
4
00;
font-weight:
7
00;
word-wrap: break-word;
"
><br
/></span
><br
/>
23 Aug
<br
/></span
><span
style=
"
color: black;
font-size: 24px;
font-family: Montserrat;
font-weight: 700;
word-wrap: break-word;
"
>
Colony PCR (Attempt 2)
</span
><span
style=
"
color: black;
...
...
@@ -239,12 +359,17 @@
font-weight: 400;
word-wrap: break-word;
"
>
Review of other work on predictive design of ribosome binding sites
- thermodynamic models, kinetic models, stochastic models, machine
learning models
<br
/>
Installing the ViennaRNA library for RNA
sequence analysis using Python
<br
/>
Downloading a dataset of 1014
RBS sequences with experimentally determined expression levels from
Reis and Salis (2020)
<br
/></span
>
(unamplified sequences) : Reduced annealing temperature to 54, 53
<br
/></span
><span
style=
"
color: black;
font-size: 24px;
font-family: Montserrat;
font-weight: 700;
word-wrap: break-word;
"
>
Gel Check
</span
><span
style=
"
color: black;
...
...
@@ -253,7 +378,8 @@
font-weight: 400;
word-wrap: break-word;
"
><br
/></span
>
:
<br
/></span
><span
style=
"
color: black;
...
...
@@ -262,12 +388,7 @@
font-weight: 400;
word-wrap: break-word;
"
>
Defining the interaction between the rRNA and mRNA so as to have
minimum hybridisation energy
<br
/>
Including the effect of spacing
between the SD sequence and the start codon
<br
/>
Defining the
interaction between the rRNA and the ribosomal S1 protein
<br
/>
Accounting
for the transient regional folding of the mRNA that can hinder RBS
accessibility
<br
/></span
>
9, 10, 15: Strong bands
<br
/>
7, 11: weak bands
<br
/></span
><span
style=
"
color: black;
...
...
@@ -282,134 +403,58 @@
color: black;
font-size: 24px;
font-family: Montserrat;
font-weight:
4
00;
font-weight:
7
00;
word-wrap: break-word;
"
>
Including the presence of standby sites upstream of the SD sequence
to which the ribosome can bind if the SD sequence is transiently
folded
<br
/>
Accounting for the effect of various start codons on
translation efficiency by using data from Hecht et al. (2017)
<br
/>
Training
and testing different machine learning models to see which works
best for the data at hand
</span
>
</div>
</div>
<div
class=
"Group43"
style=
"
width: 1201px;
height: 484px;
left: 128px;
top: 1386px;
position: absolute;
"
>
<div
class=
"Group41"
style=
"
width: 200px;
height: 478px;
left: 0px;
top: 0px;
position: absolute;
"
>
<div
class=
"JulyWeek1"
>
24 Aug
<br
/></span
><span
style=
"
width: 200px
;
height: 156
px;
left: 0px
;
top: 0px
;
position: absolute
;
color: black
;
font-size: 24
px;
font-family: Montserrat
;
font-weight: 400
;
word-wrap: break-word
;
"
>
<span
style=
"
color: #221023;
font-size: 36px;
font-family: Montserrat;
font-weight: 700;
word-wrap: break-word;
"
>
July
<br
/></span
><span
style=
"
color: #221023;
font-size: 24px;
font-family: Montserrat;
font-weight: 400;
word-wrap: break-word;
"
><br
/></span
><span
style=
"
color: #221023;
font-size: 24px;
font-family: Montserrat;
font-weight: 700;
word-wrap: break-word;
"
>
Week 1
</span
>
</div>
<div
class=
"Week2"
</span
><span
style=
"
width: 200px;
height: 156px;
left: 0px;
top: 219px;
position: absolute;
color: #221023;
color: black;
font-size: 24px;
font-family: Montserrat;
font-weight: 700;
word-wrap: break-word;
"
>
PCR Attempt 3
</span
><span
style=
"
color: black;
font-size: 24px;
font-family: Montserrat;
font-weight: 400;
word-wrap: break-word;
"
>
Week 2
</div>
<div
class=
"Week3"
(All Samples) : 1uL pure plasmid taken as template
<br
/></span
><span
style=
"
width: 200px;
height: 156px;
left: 0px;
top: 279px;
position: absolute;
color: #221023;
color: black;
font-size: 24px;
font-family: Montserrat;
font-weight: 700;
word-wrap: break-word;
"
>
Week 3
</div>
<div
class=
"Week4"
><br
/>
25 Aug
<br
/></span
><span
style=
"
width: 200px;
height: 23px;
left: 0px;
top: 455px;
position: absolute;
color: #221023;
color: black;
font-size: 24px;
font-family: Montserrat;
font-weight: 700;
word-wrap: break-word;
"
>
Week 4
</div>
</div>
<div
class=
"DownloadingADatasetOf16779RbsSequencesCharacterisedByFlowseqFromReisAndSalis2020AndTestingTheModelOnItReadingUpOnDifferentOptimisationAlgorithmsThatCanBeUsedToMutateAnRbsSequenceInSilicoInOrderToAchieveADesiredExpressionLevelSimulatedAnnealingAndGradientDescentOptimisersWereCreatedAndTestedOptimiserPerformanceWasTestedByVaryingItsIntrinsicParametersWorkIsOngoingAPartiallyWorkingGeneticAlgorithmWasCreatedWorkIsYetToBeFinishedOnThisTheTwoWorkingOptimisersWereUsedToGenerateRbsSequencesWithVaryingExpressionLevelsTheseSequencesWillBeUsedInTheWetLabUsingGfpExpressingLactococcusLactisToSeeIfTheOptimisationWasSuccessfulAllIndiaIgemMeet"
style=
"width: 1079px; left: 122px; top: 72px; position: absolute"
>
<span
>
Gel Check
</span
><span
style=
"
color: black;
font-size: 24px;
...
...
@@ -417,11 +462,8 @@
font-weight: 400;
word-wrap: break-word;
"
>
Downloading a dataset of 16779 RBS sequences characterised by
FlowSeq from Reis and Salis (2020) and testing the model on it
<br
/>
Reading
up on different optimisation algorithms that can be used to "mutate"
an RBS sequence in silico in order to achieve a desired expression
level
<br
/></span
>
: 4, 5, 7, 8, 9 14 Not amplified
<br
/></span
><span
style=
"
color: black;
...
...
@@ -436,20 +478,19 @@
color: black;
font-size: 24px;
font-family: Montserrat;
font-weight:
4
00;
font-weight:
7
00;
word-wrap: break-word;
"
>
Simulated annealing and gradient descent optimisers were created
and tested
<br
/></span
>
27 Aug
<br
/></span
><span
style=
"
color: black;
font-size: 24px;
font-family: Montserrat;
font-weight:
4
00;
font-weight:
7
00;
word-wrap: break-word;
"
>
<br
/>
</span
>
PCR Attempt 4
</span
><span
style=
"
color: black;
...
...
@@ -458,22 +499,17 @@
font-weight: 400;
word-wrap: break-word;
"
>
Optimiser performance was tested by varying its intrinsic
parameters, work is ongoing
<br
/>
A partially working genetic
algorithm was created, work is yet to be finished on this
<br
/>
The
two working optimisers were used to generate RBS sequences with
varying expression levels. These sequences will be used in the wet
lab using GFP-expressing Lactococcus lactis to see if the
optimisation was successful.
<br
/></span
>
(Unamplified samples) : Reduced annealing temperature to 53, 52
<br
/></span
><span
style=
"
color: black;
font-size: 24px;
font-family: Montserrat;
font-weight:
4
00;
font-weight:
7
00;
word-wrap: break-word;
"
>
<br
/>
</span
>
Gel Check
</span
><span
style=
"
color: black;
...
...
@@ -482,126 +518,54 @@
font-weight: 400;
word-wrap: break-word;
"
>
All India iGEM Meet
</span
>
</div>
</div>
<div
class=
"Group43"
style=
"
width: 1201px;
height: 397px;
left: 128px;
top: 1958px;
position: absolute;
"
>
<div
class=
"Group41"
style=
"
width: 200px;
height: 391px;
left: 0px;
top: 0px;
position: absolute;
"
>
<div
class=
"AugustWeek1"
: only 9 Amplified
<br
/></span
><span
style=
"
width: 200px
;
height: 156
px;
left: 0px
;
top: 0px
;
position: absolute
;
color: black
;
font-size: 24
px;
font-family: Montserrat
;
font-weight: 400
;
word-wrap: break-word
;
"
>
<span
style=
"
color: #221023;
font-size: 36px;
font-family: Montserrat;
font-weight: 700;
word-wrap: break-word;
"
>
August
<br
/></span
><span
style=
"
color: #221023;
font-size: 24px;
font-family: Montserrat;
font-weight: 400;
word-wrap: break-word;
"
><br
/></span
><span
style=
"
color: #221023;
font-size: 24px;
font-family: Montserrat;
font-weight: 700;
word-wrap: break-word;
"
>
Week 1
</span
>
</div>
<div
class=
"Week2"
><br
/></span
><span
style=
"
width: 200px;
height: 156px;
left: 0px;
top: 132px;
position: absolute;
color: #221023;
color: black;
font-size: 24px;
font-family: Montserrat;
font-weight: 700;
word-wrap: break-word;
"
>
Week 2
</div>
<div
class=
"Week3"
>
28 Aug
<br
/></span
><span
style=
"
width: 200px;
height: 112px;
left: 0px;
top: 279px;
position: absolute;
color: #221023;
color: black;
font-size: 24px;
font-family: Montserrat;
font-weight: 700;
word-wrap: break-word;
"
>
Week 3
</div>
<div
class=
"Week4"
>
PCR Attempt 5
</span
><span
style=
"
color: black;
font-size: 24px;
font-family: Montserrat;
font-weight: 400;
word-wrap: break-word;
"
>
(Unamplified samples) : Touchdown PCR 56-52C
<br
/></span
><span
style=
"
width: 200px;
height: 23px;
left: 0px;
top: 339px;
position: absolute;
color: #221023;
color: black;
font-size: 24px;
font-family: Montserrat;
font-weight: 700;
word-wrap: break-word;
"
>
Week 4
</div>
</div>
<div
class=
"GeneticAlgorithmCompletedFoundToHaveTheBestPerformanceAttemptedToCombineDifferentAspectsToTheOptimizationAlgorithmsStartedWorkOnAddingNewParametersToTheModelToImproveAccuracyOfPredictionsLiteratureReviewOnRnaFoldingKineticsAndRnaDegradationWasInitiatedLiteratureReviewContinuedStartingWorkingWithTheKinfoldLibraryOfViennarnaToFindTheFoldingTimeThereWasANeedToIntegrateCommandLineOperationsWithPythonCode"
style=
"width: 1079px; left: 122px; top: 72px; position: absolute"
>
<span
>
Gel Check
</span
><span
style=
"
color: black;
font-size: 24px;
...
...
@@ -609,7 +573,8 @@
font-weight: 400;
word-wrap: break-word;
"
>
Genetic algorithm completed. Found to have the best performance.
<br
/></span
>
: 4, 5 Unamplified
<br
/></span
><span
style=
"
color: black;
...
...
@@ -624,13 +589,19 @@
color: black;
font-size: 24px;
font-family: Montserrat;
font-weight: 400;
font-weight: 700;
word-wrap: break-word;
"
>
30 Aug
<br
/></span
><span
style=
"
color: black;
font-size: 24px;
font-family: Montserrat;
font-weight: 700;
word-wrap: break-word;
"
>
Attempted to combine different aspects to the optimization
algorithms.
<br
/>
Started work on adding new parameters to the model
to improve accuracy of predictions. Literature review on RNA folding
kinetics and RNA degradation was initiated.
<br
/></span
>
PCR Attempt 6
</span
><span
style=
"
color: black;
...
...
@@ -639,7 +610,18 @@
font-weight: 400;
word-wrap: break-word;
"
><br
/></span
>
(Unamplified samples) : 50uL reaction separated into 2 tubes of 25uL
<br
/></span
><span
style=
"
color: black;
font-size: 24px;
font-family: Montserrat;
font-weight: 700;
word-wrap: break-word;
"
>
Gel Check
</span
><span
style=
"
color: black;
...
...
@@ -648,7 +630,8 @@
font-weight: 400;
word-wrap: break-word;
"
>
Literature review continued
<br
/></span
>
: both fragments (4, 5) amplified
<br
/></span
><span
style=
"
color: black;
...
...
@@ -658,6 +641,15 @@
word-wrap: break-word;
"
><br
/></span
><span
style=
"
color: black;
font-size: 24px;
font-family: Montserrat;
font-weight: 700;
word-wrap: break-word;
"
>
All fragments have been amplified!
<br
/></span
><span
style=
"
color: black;
...
...
@@ -666,673 +658,774 @@
font-weight: 400;
word-wrap: break-word;
"
>
Starting working with the Kinfold library of ViennaRNA to find the
folding time. There was a need to integrate command line operations
with Python code.
</span
>
</div>
</div>
<div
class=
"Group46Default"
style=
"
width: 1201px;
height: 362px;
left: 128px;
top: 2443px;
position: absolute;
justify-content: center;
align-items: center;
display: inline-flex;
"
>
<div
class=
"Group43"
style=
"width: 1201px; height: 362px; position: relative"
>
<div
class=
"Group41"
><br
/></span
><span
style=
"
width: 226px
;
height: 362
px;
left: 0px
;
top: 0px
;
position: absolute
;
color: black
;
font-size: 24
px;
font-family: Montserrat
;
font-weight: 700
;
word-wrap: break-word
;
"
>
<div
class=
"SeptemberWeek1"
style=
"
width: 226px;
height: 156px;
left: 0px;
top: 0px;
position: absolute;
"
>
<span
style=
"
color: #221023;
font-size: 36px;
font-family: Montserrat;
font-weight: 700;
word-wrap: break-word;
"
>
September
<br
/></span
><span
style=
"
color: #221023;
font-size: 24px;
font-family: Montserrat;
font-weight: 400;
word-wrap: break-word;
"
><br
/></span
><span
style=
"
color: #221023;
font-size: 24px;
font-family: Montserrat;
font-weight: 700;
word-wrap: break-word;
"
>
Week 1
</span
>
</div>
<div
class=
"Week2"
style=
"
width: 200px;
height: 156px;
left: 0px;
top: 161px;
position: absolute;
color: #221023;
font-size: 24px;
font-family: Montserrat;
font-weight: 700;
word-wrap: break-word;
"
>
Week 2
</div>
<div
class=
"Week3"
style=
"
width: 200px;
height: 112px;
left: 0px;
top: 239px;
position: absolute;
color: #221023;
font-size: 24px;
font-family: Montserrat;
font-weight: 700;
word-wrap: break-word;
"
>
Week 3
</div>
<div
class=
"Week4"
style=
"
width: 200px;
height: 23px;
left: 0px;
top: 339px;
position: absolute;
color: #221023;
font-size: 24px;
font-family: Montserrat;
font-weight: 700;
word-wrap: break-word;
"
>
Week 4
</div>
</div>
<div
class=
"ContinuedAttemptToGetKinfoldToWorkLiteratureReviewConcludedKinfoldFinallyStartedWorkingPrototypeForDegradationModelBuilt"
style=
"width: 1079px; left: 122px; top: 72px; position: absolute"
>
<span
style=
"
color: black;
font-size: 24px;
font-family: Montserrat;
font-weight: 400;
word-wrap: break-word;
"
>
Continued attempt to get Kinfold to work
<br
/>
Literature review
concluded
<br
/></span
><span
style=
"
color: black;
font-size: 24px;
font-family: Montserrat;
font-weight: 400;
word-wrap: break-word;
"
><br
/></span
><span
style=
"
color: black;
font-size: 24px;
font-family: Montserrat;
font-weight: 400;
word-wrap: break-word;
"
>
Kinfold finally started working
<br
/>
Prototype for degradation
model built
</span
>
</div>
>
31 Aug
<br
/></span
><span
style=
"
color: black;
font-size: 24px;
font-family: Montserrat;
font-weight: 400;
word-wrap: break-word;
"
>
0.4uL Dpn1 added to all samples
<br
/></span
><span
style=
"
color: black;
font-size: 24px;
font-family: Montserrat;
font-weight: 700;
word-wrap: break-word;
"
>
Gel Extraction
</span
><span
style=
"
color: black;
font-size: 24px;
font-family: Montserrat;
font-weight: 400;
word-wrap: break-word;
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>
: (Qiagen Kit protocol)
<br
/></span
><span
style=
"
color: black;
font-size: 24px;
font-family: Montserrat;
font-weight: 400;
word-wrap: break-word;
"
>
Plasmid conc :
<br
/></span
><span
style=
"
color: black;
font-size: 24px;
font-family: Montserrat;
font-weight: 400;
word-wrap: break-word;
"
>
3,4,5,6,7,8,9,10 - around 10ng/uL.
<br
/>
11,12,13,14,15,16 - around
30-40 ng/uL
<br
/>
5 - 2ng/ul
</span
>
</div>
<div
style=
"width: 1079px; left: 0px; top: 72px; position: absolute"
>
<span
style=
"
color: black;
font-size: 24px;
font-family: Montserrat;
font-weight: 700;
word-wrap: break-word;
"
>
18 Aug
<br
/></span
><span
style=
"
color: black;
font-size: 24px;
font-family: Montserrat;
font-weight: 400;
word-wrap: break-word;
"
>
Randomized primers for both UTR regions received.
<br
/>
Lyophilized
primers made to 100uM conc and resuspended. Dilutions of 10uM are
made.
</span
>
</div>
<div
style=
"
width: 801px;
left: 160px;
top: 1002px;
position: absolute;
color: black;
font-size: 20px;
font-family: Montserrat;
font-weight: 400;
word-wrap: break-word;
"
>
Sequences are referred to by serial numbers
<br
/><br
/>
Optimization
Algorithms used to generate the sequence:
<br
/>
SA - Simulated
Annealing
<br
/>
GD - Gradient Descent
<br
/>
GA - Genetic Algorithm
<br
/><br
/>
H,
L = High, Low Expressing
</div>
</div>
<div
style=
"
width: 1079px;
height: 1981px;
left: 181px;
top: 3068px;
position: absolute;
"
>
<div
style=
"
width: 357px;
height: 156px;
left: 0px;
top: 0px;
position: absolute;
color: #221023;
font-size: 36px;
font-family: Montserrat;
font-weight: 700;
word-wrap: break-word;
"
>
September
</div>
<div
style=
"width: 1079px; left: 0px; top: 72px; position: absolute"
>
<span
style=
"
color: black;
font-size: 24px;
font-family: Montserrat;
font-weight: 700;
word-wrap: break-word;
"
>
2 Sept
<br
/></span
><span
style=
"
color: black;
font-size: 24px;
font-family: Montserrat;
font-weight: 400;
word-wrap: break-word;
"
>
Transformation + Plating
<br
/></span
><span
style=
"
color: black;
font-size: 24px;
font-family: Montserrat;
font-weight: 400;
word-wrap: break-word;
"
>
Electroporation was done for all the samples in
</span
><span
style=
"
color: black;
font-size: 24px;
font-family: Montserrat;
font-style: italic;
font-weight: 400;
word-wrap: break-word;
"
>
L. lactis
</span
><span
style=
"
color: black;
font-size: 24px;
font-family: Montserrat;
font-weight: 400;
word-wrap: break-word;
"
>
competent cells.
<br
/>
Samples were incubated for 1hr after which
they were spread plated and incubated for 24 hours.
<br
/></span
><span
style=
"
color: black;
font-size: 24px;
font-family: Montserrat;
font-weight: 400;
word-wrap: break-word;
"
><br
/></span
><span
style=
"
color: black;
font-size: 24px;
font-family: Montserrat;
font-weight: 700;
word-wrap: break-word;
"
>
3 Sept
<br
/></span
><span
style=
"
color: black;
font-size: 24px;
font-family: Montserrat;
font-weight: 400;
word-wrap: break-word;
"
>
Colonies in all plates except samples 4 and 14. 10 colonies were
observed in plate 3.
<br
/>
Colonies were picked and cultured
overnight. (2 cultures from each non-randomized sample, all 10
cultures of each colony from the randomized sample 3.
<br
/></span
><span
style=
"
color: black;
font-size: 24px;
font-family: Montserrat;
font-weight: 700;
word-wrap: break-word;
"
><br
/>
4 Sept
<br
/></span
><span
style=
"
color: black;
font-size: 24px;
font-family: Montserrat;
font-weight: 400;
word-wrap: break-word;
"
>
Glycerol added and strains stored in -80C
<br
/></span
><span
style=
"
color: black;
font-size: 24px;
font-family: Montserrat;
font-weight: 700;
word-wrap: break-word;
"
><br
/>
5 Sept
<br
/></span
><span
style=
"
color: black;
font-size: 24px;
font-family: Montserrat;
font-weight: 400;
word-wrap: break-word;
"
>
Transformation and plating (Attempt 2) (Samples 3,4,14) (Working to
obtain more strains of randomized sample 3)
<br
/></span
><span
style=
"
color: black;
font-size: 24px;
font-family: Montserrat;
font-weight: 400;
word-wrap: break-word;
"
><br
/></span
><span
style=
"
color: black;
font-size: 24px;
font-family: Montserrat;
font-weight: 700;
word-wrap: break-word;
"
>
6 Sept
<br
/></span
><span
style=
"
color: black;
font-size: 24px;
font-family: Montserrat;
font-weight: 400;
word-wrap: break-word;
"
>
No colonies observed
<br
/></span
><span
style=
"
color: black;
font-size: 24px;
font-family: Montserrat;
font-weight: 400;
word-wrap: break-word;
"
><br
/></span
><span
style=
"
color: black;
font-size: 24px;
font-family: Montserrat;
font-weight: 700;
word-wrap: break-word;
"
>
8 Sept
<br
/></span
><span
style=
"
color: black;
font-size: 24px;
font-family: Montserrat;
font-weight: 400;
word-wrap: break-word;
"
>
Transformation and plating (Attempt 4) (Samples 3,4,14)
<br
/></span
><span
style=
"
color: black;
font-size: 24px;
font-family: Montserrat;
font-weight: 400;
word-wrap: break-word;
"
><br
/></span
><span
style=
"
color: black;
font-size: 24px;
font-family: Montserrat;
font-weight: 700;
word-wrap: break-word;
"
>
9 Sept
<br
/></span
><span
style=
"
color: black;
font-size: 24px;
font-family: Montserrat;
font-weight: 400;
word-wrap: break-word;
"
>
No colonies observed
<br
/></span
><span
style=
"
color: black;
font-size: 24px;
font-family: Montserrat;
font-weight: 400;
word-wrap: break-word;
"
><br
/></span
><span
style=
"
color: black;
font-size: 24px;
font-family: Montserrat;
font-weight: 700;
word-wrap: break-word;
"
>
11 Sept
<br
/></span
><span
style=
"
color: black;
font-size: 24px;
font-family: Montserrat;
font-weight: 400;
word-wrap: break-word;
"
>
Transformation and plating (Attempt 5) (Samples 3,4,14) (Slightly
reduced antibiotic concentration)
<br
/></span
><span
style=
"
color: black;
font-size: 24px;
font-family: Montserrat;
font-weight: 400;
word-wrap: break-word;
"
><br
/></span
><span
style=
"
color: black;
font-size: 24px;
font-family: Montserrat;
font-weight: 700;
word-wrap: break-word;
"
>
12 Sept
<br
/></span
><span
style=
"
color: black;
font-size: 24px;
font-family: Montserrat;
font-weight: 400;
word-wrap: break-word;
"
>
Golden Gate Assembly set up from left over plasmid for 3,4,14
<br
/></span
><span
style=
"
color: black;
font-size: 24px;
font-family: Montserrat;
font-weight: 400;
word-wrap: break-word;
"
><br
/></span
><span
style=
"
color: black;
font-size: 24px;
font-family: Montserrat;
font-weight: 700;
word-wrap: break-word;
"
>
14 Sept
<br
/></span
><span
style=
"
color: black;
font-size: 24px;
font-family: Montserrat;
font-weight: 400;
word-wrap: break-word;
"
>
Transformation and plating (Attempt 6) (Samples 3,4,14)
<br
/></span
><span
style=
"
color: black;
font-size: 24px;
font-family: Montserrat;
font-weight: 400;
word-wrap: break-word;
"
><br
/></span
><span
style=
"
color: black;
font-size: 24px;
font-family: Montserrat;
font-weight: 700;
word-wrap: break-word;
"
>
15 Sept
<br
/></span
><span
style=
"
color: black;
font-size: 24px;
font-family: Montserrat;
font-weight: 400;
word-wrap: break-word;
"
>
1 colony for sample 3. None for 4 and 14.
<br
/></span
><span
style=
"
color: black;
font-size: 24px;
font-family: Montserrat;
font-weight: 400;
word-wrap: break-word;
"
><br
/></span
><span
style=
"
color: black;
font-size: 24px;
font-family: Montserrat;
font-weight: 700;
word-wrap: break-word;
"
>
17 Sept
<br
/></span
><span
style=
"
color: black;
font-size: 24px;
font-family: Montserrat;
font-weight: 400;
word-wrap: break-word;
"
>
Transformation and plating (Attempt 7) (Samples 3,4,14)
<br
/></span
><span
style=
"
color: black;
font-size: 24px;
font-family: Montserrat;
font-weight: 400;
word-wrap: break-word;
"
><br
/></span
><span
style=
"
color: black;
font-size: 24px;
font-family: Montserrat;
font-weight: 700;
word-wrap: break-word;
"
>
18 Sept
<br
/></span
><span
style=
"
color: black;
font-size: 24px;
font-family: Montserrat;
font-weight: 400;
word-wrap: break-word;
"
>
2 colonies in sample 3. None for 4 and 14.
<br
/></span
><span
style=
"
color: black;
font-size: 24px;
font-family: Montserrat;
font-weight: 400;
word-wrap: break-word;
"
><br
/></span
><span
style=
"
color: black;
font-size: 24px;
font-family: Montserrat;
font-weight: 700;
word-wrap: break-word;
"
>
21 Sept
<br
/></span
><span
style=
"
color: black;
font-size: 24px;
font-family: Montserrat;
font-weight: 400;
word-wrap: break-word;
"
>
All strains (13 variants from randomized sample 3, 11 model
generated sequences) streaked on agar plates
<br
/></span
><span
style=
"
color: black;
font-size: 24px;
font-family: Montserrat;
font-weight: 400;
word-wrap: break-word;
"
><br
/></span
><span
style=
"
color: black;
font-size: 24px;
font-family: Montserrat;
font-weight: 700;
word-wrap: break-word;
"
>
22 Sept
<br
/></span
><span
style=
"
color: black;
font-size: 24px;
font-family: Montserrat;
font-weight: 400;
word-wrap: break-word;
"
>
Colonies picked and Overnight 1ml cultures made and incubated for
14 hrs
<br
/></span
><span
style=
"
color: black;
font-size: 24px;
font-family: Montserrat;
font-weight: 400;
word-wrap: break-word;
"
><br
/></span
><span
style=
"
color: black;
font-size: 24px;
font-family: Montserrat;
font-weight: 700;
word-wrap: break-word;
"
>
23 Sept
<br
/></span
><span
style=
"
color: black;
font-size: 24px;
font-family: Montserrat;
font-weight: 400;
word-wrap: break-word;
"
>
Fluorescence Assay
<br
/></span
><span
style=
"
color: black;
font-size: 24px;
font-family: Montserrat;
font-weight: 700;
word-wrap: break-word;
"
>
First batch of results obtained.
<br
/></span
><span
style=
"
color: black;
font-size: 24px;
font-family: Montserrat;
font-weight: 400;
word-wrap: break-word;
"
><br
/></span
><span
style=
"
color: black;
font-size: 24px;
font-family: Montserrat;
font-weight: 700;
word-wrap: break-word;
"
>
25 Sept
<br
/></span
><span
style=
"
color: black;
font-size: 24px;
font-family: Montserrat;
font-weight: 400;
word-wrap: break-word;
"
>
Colonies picked from streaked plates and cultured overnight.
<br
/></span
><span
style=
"
color: black;
font-size: 24px;
font-family: Montserrat;
font-weight: 400;
word-wrap: break-word;
"
><br
/></span
><span
style=
"
color: black;
font-size: 24px;
font-family: Montserrat;
font-weight: 700;
word-wrap: break-word;
"
>
26 Sept
<br
/></span
><span
style=
"
color: black;
font-size: 24px;
font-family: Montserrat;
font-weight: 400;
word-wrap: break-word;
"
>
Plasmid isolation (Qiagen Kit protocol)
<br
/>
Plasmid concentration
obtained were very poor.
<br
/></span>
</div>
</div>
</div>
//---------------------------------------------------------------
//Wetlab Notebook
<div
class=
"wet-lab-notebook-yXv"
id=
"400:7"
>
<div
class=
"rectangle-7-ZW8"
id=
"400:8"
></div>
<div
class=
"auto-group-7tjh-q6G"
id=
"LWGbMeqr3bqpVb9LaJ7tJH"
>
Lab Notebook
</div>
<div
class=
"feature-engineering-RKN"
id=
"400:11"
>
Wet Lab
</div>
<div
class=
"auto-group-h1nu-V4L"
id=
"LWGbUyxy1yythvL3pBH1Nu"
>
<div
class=
"group-43-PQc"
id=
"400:30"
>
<div
class=
"auto-group-z6ff-hgC"
id=
"LWGbfEAtvRerghbgN6z6fF"
>
<p
class=
"august-zvC"
id=
"400:32"
>
August
<br/>
</p>
<p
class=
"aug-randomized-primers-for-both-utr-regions-received-lyophilized-primers-made-to-100um-conc-and-resuspended-dilutions-of-10um-are-made-FLL"
id=
"539:51"
>
<span
class=
"aug-randomized-primers-for-both-utr-regions-received-lyophilized-primers-made-to-100um-conc-and-resuspended-dilutions-of-10um-are-made-FLL-sub-0"
>
18 Aug
<br/>
</span>
<span
class=
"aug-randomized-primers-for-both-utr-regions-received-lyophilized-primers-made-to-100um-conc-and-resuspended-dilutions-of-10um-are-made-FLL-sub-1"
>
Randomized primers for both UTR regions received.
<br/>
Lyophilized primers made to 100uM conc and resuspended. Dilutions of 10uM are made.
</span>
</p>
</div>
<div
class=
"screenshot-2023-10-11-at-910-1-8wr"
id=
"539:53"
></div>
<div
class=
"auto-group-gsz1-P76"
id=
"LWGbjthnp875upXx4aGSz1"
>
<p
class=
"aug-colony-pcr-attempt-1-template-solution-was-made-by-picking-colonies-and-dispersing-in-aq-medium-21-aug-gel-check-all-pcr-products-ran-on-agarose-gel-6-8-13-16-amplified-23-aug-colony-pcr-attempt-2-unamplified-sequences-reduced-annealing-temperature-to-54-53-gel-check-9-10-15-strong-bands-7-11-weak-bands-24-aug-pcr-attempt-3-all-samples-1ul-pure-plasmid-taken-as-template-25-aug-gel-check-4-5-7-8-9-14-not-amplified-27-aug-pcr-attempt-4-unamplified-samples-reduced-annealing-temperature-to-53-52-gel-check-only-9-amplified-28-aug-pcr-attempt-5-unamplified-samples-touchdown-pcr-56-52c-gel-check-4-5-unamplified-30-aug-pcr-attempt-6-unamplified-samples-50ul-reaction-separated-into-2-tubes-of-25ul-gel-check-both-fragments-4-5-amplified-all-fragments-have-been-amplified-31-aug-04ul-dpn1-added-to-all-samples-gel-extraction-qiagen-kit-protocol-plasmid-conc-345678910-around-10ng-ul-111213141516-around-30-40-ng-ul-5-2ng-ul-7Yt"
id=
"400:36"
>
<span
class=
"aug-colony-pcr-attempt-1-template-solution-was-made-by-picking-colonies-and-dispersing-in-aq-medium-21-aug-gel-check-all-pcr-products-ran-on-agarose-gel-6-8-13-16-amplified-23-aug-colony-pcr-attempt-2-unamplified-sequences-reduced-annealing-temperature-to-54-53-gel-check-9-10-15-strong-bands-7-11-weak-bands-24-aug-pcr-attempt-3-all-samples-1ul-pure-plasmid-taken-as-template-25-aug-gel-check-4-5-7-8-9-14-not-amplified-27-aug-pcr-attempt-4-unamplified-samples-reduced-annealing-temperature-to-53-52-gel-check-only-9-amplified-28-aug-pcr-attempt-5-unamplified-samples-touchdown-pcr-56-52c-gel-check-4-5-unamplified-30-aug-pcr-attempt-6-unamplified-samples-50ul-reaction-separated-into-2-tubes-of-25ul-gel-check-both-fragments-4-5-amplified-all-fragments-have-been-amplified-31-aug-04ul-dpn1-added-to-all-samples-gel-extraction-qiagen-kit-protocol-plasmid-conc-345678910-around-10ng-ul-111213141516-around-30-40-ng-ul-5-2ng-ul-7Yt-sub-0"
>
<br/>
<br/>
</span>
<span
class=
"aug-colony-pcr-attempt-1-template-solution-was-made-by-picking-colonies-and-dispersing-in-aq-medium-21-aug-gel-check-all-pcr-products-ran-on-agarose-gel-6-8-13-16-amplified-23-aug-colony-pcr-attempt-2-unamplified-sequences-reduced-annealing-temperature-to-54-53-gel-check-9-10-15-strong-bands-7-11-weak-bands-24-aug-pcr-attempt-3-all-samples-1ul-pure-plasmid-taken-as-template-25-aug-gel-check-4-5-7-8-9-14-not-amplified-27-aug-pcr-attempt-4-unamplified-samples-reduced-annealing-temperature-to-53-52-gel-check-only-9-amplified-28-aug-pcr-attempt-5-unamplified-samples-touchdown-pcr-56-52c-gel-check-4-5-unamplified-30-aug-pcr-attempt-6-unamplified-samples-50ul-reaction-separated-into-2-tubes-of-25ul-gel-check-both-fragments-4-5-amplified-all-fragments-have-been-amplified-31-aug-04ul-dpn1-added-to-all-samples-gel-extraction-qiagen-kit-protocol-plasmid-conc-345678910-around-10ng-ul-111213141516-around-30-40-ng-ul-5-2ng-ul-7Yt-sub-1"
>
19 Aug
<br/>
Colony PCR (Attempt 1)
</span>
<span
class=
"aug-colony-pcr-attempt-1-template-solution-was-made-by-picking-colonies-and-dispersing-in-aq-medium-21-aug-gel-check-all-pcr-products-ran-on-agarose-gel-6-8-13-16-amplified-23-aug-colony-pcr-attempt-2-unamplified-sequences-reduced-annealing-temperature-to-54-53-gel-check-9-10-15-strong-bands-7-11-weak-bands-24-aug-pcr-attempt-3-all-samples-1ul-pure-plasmid-taken-as-template-25-aug-gel-check-4-5-7-8-9-14-not-amplified-27-aug-pcr-attempt-4-unamplified-samples-reduced-annealing-temperature-to-53-52-gel-check-only-9-amplified-28-aug-pcr-attempt-5-unamplified-samples-touchdown-pcr-56-52c-gel-check-4-5-unamplified-30-aug-pcr-attempt-6-unamplified-samples-50ul-reaction-separated-into-2-tubes-of-25ul-gel-check-both-fragments-4-5-amplified-all-fragments-have-been-amplified-31-aug-04ul-dpn1-added-to-all-samples-gel-extraction-qiagen-kit-protocol-plasmid-conc-345678910-around-10ng-ul-111213141516-around-30-40-ng-ul-5-2ng-ul-7Yt-sub-2"
>
: Template solution was made by picking colonies and dispersing in aq medium.
<br/>
</span>
<span
class=
"aug-colony-pcr-attempt-1-template-solution-was-made-by-picking-colonies-and-dispersing-in-aq-medium-21-aug-gel-check-all-pcr-products-ran-on-agarose-gel-6-8-13-16-amplified-23-aug-colony-pcr-attempt-2-unamplified-sequences-reduced-annealing-temperature-to-54-53-gel-check-9-10-15-strong-bands-7-11-weak-bands-24-aug-pcr-attempt-3-all-samples-1ul-pure-plasmid-taken-as-template-25-aug-gel-check-4-5-7-8-9-14-not-amplified-27-aug-pcr-attempt-4-unamplified-samples-reduced-annealing-temperature-to-53-52-gel-check-only-9-amplified-28-aug-pcr-attempt-5-unamplified-samples-touchdown-pcr-56-52c-gel-check-4-5-unamplified-30-aug-pcr-attempt-6-unamplified-samples-50ul-reaction-separated-into-2-tubes-of-25ul-gel-check-both-fragments-4-5-amplified-all-fragments-have-been-amplified-31-aug-04ul-dpn1-added-to-all-samples-gel-extraction-qiagen-kit-protocol-plasmid-conc-345678910-around-10ng-ul-111213141516-around-30-40-ng-ul-5-2ng-ul-7Yt-sub-3"
>
<br/>
21 Aug
<br/>
Gel Check
</span>
<span
class=
"aug-colony-pcr-attempt-1-template-solution-was-made-by-picking-colonies-and-dispersing-in-aq-medium-21-aug-gel-check-all-pcr-products-ran-on-agarose-gel-6-8-13-16-amplified-23-aug-colony-pcr-attempt-2-unamplified-sequences-reduced-annealing-temperature-to-54-53-gel-check-9-10-15-strong-bands-7-11-weak-bands-24-aug-pcr-attempt-3-all-samples-1ul-pure-plasmid-taken-as-template-25-aug-gel-check-4-5-7-8-9-14-not-amplified-27-aug-pcr-attempt-4-unamplified-samples-reduced-annealing-temperature-to-53-52-gel-check-only-9-amplified-28-aug-pcr-attempt-5-unamplified-samples-touchdown-pcr-56-52c-gel-check-4-5-unamplified-30-aug-pcr-attempt-6-unamplified-samples-50ul-reaction-separated-into-2-tubes-of-25ul-gel-check-both-fragments-4-5-amplified-all-fragments-have-been-amplified-31-aug-04ul-dpn1-added-to-all-samples-gel-extraction-qiagen-kit-protocol-plasmid-conc-345678910-around-10ng-ul-111213141516-around-30-40-ng-ul-5-2ng-ul-7Yt-sub-4"
>
: All PCR products ran on agarose gel; 6, 8, 13, 16 amplified
<br/>
</span>
<span
class=
"aug-colony-pcr-attempt-1-template-solution-was-made-by-picking-colonies-and-dispersing-in-aq-medium-21-aug-gel-check-all-pcr-products-ran-on-agarose-gel-6-8-13-16-amplified-23-aug-colony-pcr-attempt-2-unamplified-sequences-reduced-annealing-temperature-to-54-53-gel-check-9-10-15-strong-bands-7-11-weak-bands-24-aug-pcr-attempt-3-all-samples-1ul-pure-plasmid-taken-as-template-25-aug-gel-check-4-5-7-8-9-14-not-amplified-27-aug-pcr-attempt-4-unamplified-samples-reduced-annealing-temperature-to-53-52-gel-check-only-9-amplified-28-aug-pcr-attempt-5-unamplified-samples-touchdown-pcr-56-52c-gel-check-4-5-unamplified-30-aug-pcr-attempt-6-unamplified-samples-50ul-reaction-separated-into-2-tubes-of-25ul-gel-check-both-fragments-4-5-amplified-all-fragments-have-been-amplified-31-aug-04ul-dpn1-added-to-all-samples-gel-extraction-qiagen-kit-protocol-plasmid-conc-345678910-around-10ng-ul-111213141516-around-30-40-ng-ul-5-2ng-ul-7Yt-sub-5"
>
<br/>
23 Aug
<br/>
Colony PCR (Attempt 2)
</span>
<span
class=
"aug-colony-pcr-attempt-1-template-solution-was-made-by-picking-colonies-and-dispersing-in-aq-medium-21-aug-gel-check-all-pcr-products-ran-on-agarose-gel-6-8-13-16-amplified-23-aug-colony-pcr-attempt-2-unamplified-sequences-reduced-annealing-temperature-to-54-53-gel-check-9-10-15-strong-bands-7-11-weak-bands-24-aug-pcr-attempt-3-all-samples-1ul-pure-plasmid-taken-as-template-25-aug-gel-check-4-5-7-8-9-14-not-amplified-27-aug-pcr-attempt-4-unamplified-samples-reduced-annealing-temperature-to-53-52-gel-check-only-9-amplified-28-aug-pcr-attempt-5-unamplified-samples-touchdown-pcr-56-52c-gel-check-4-5-unamplified-30-aug-pcr-attempt-6-unamplified-samples-50ul-reaction-separated-into-2-tubes-of-25ul-gel-check-both-fragments-4-5-amplified-all-fragments-have-been-amplified-31-aug-04ul-dpn1-added-to-all-samples-gel-extraction-qiagen-kit-protocol-plasmid-conc-345678910-around-10ng-ul-111213141516-around-30-40-ng-ul-5-2ng-ul-7Yt-sub-6"
>
(unamplified sequences) : Reduced annealing temperature to 54, 53
<br/>
</span>
<span
class=
"aug-colony-pcr-attempt-1-template-solution-was-made-by-picking-colonies-and-dispersing-in-aq-medium-21-aug-gel-check-all-pcr-products-ran-on-agarose-gel-6-8-13-16-amplified-23-aug-colony-pcr-attempt-2-unamplified-sequences-reduced-annealing-temperature-to-54-53-gel-check-9-10-15-strong-bands-7-11-weak-bands-24-aug-pcr-attempt-3-all-samples-1ul-pure-plasmid-taken-as-template-25-aug-gel-check-4-5-7-8-9-14-not-amplified-27-aug-pcr-attempt-4-unamplified-samples-reduced-annealing-temperature-to-53-52-gel-check-only-9-amplified-28-aug-pcr-attempt-5-unamplified-samples-touchdown-pcr-56-52c-gel-check-4-5-unamplified-30-aug-pcr-attempt-6-unamplified-samples-50ul-reaction-separated-into-2-tubes-of-25ul-gel-check-both-fragments-4-5-amplified-all-fragments-have-been-amplified-31-aug-04ul-dpn1-added-to-all-samples-gel-extraction-qiagen-kit-protocol-plasmid-conc-345678910-around-10ng-ul-111213141516-around-30-40-ng-ul-5-2ng-ul-7Yt-sub-7"
>
Gel Check
</span>
<span
class=
"aug-colony-pcr-attempt-1-template-solution-was-made-by-picking-colonies-and-dispersing-in-aq-medium-21-aug-gel-check-all-pcr-products-ran-on-agarose-gel-6-8-13-16-amplified-23-aug-colony-pcr-attempt-2-unamplified-sequences-reduced-annealing-temperature-to-54-53-gel-check-9-10-15-strong-bands-7-11-weak-bands-24-aug-pcr-attempt-3-all-samples-1ul-pure-plasmid-taken-as-template-25-aug-gel-check-4-5-7-8-9-14-not-amplified-27-aug-pcr-attempt-4-unamplified-samples-reduced-annealing-temperature-to-53-52-gel-check-only-9-amplified-28-aug-pcr-attempt-5-unamplified-samples-touchdown-pcr-56-52c-gel-check-4-5-unamplified-30-aug-pcr-attempt-6-unamplified-samples-50ul-reaction-separated-into-2-tubes-of-25ul-gel-check-both-fragments-4-5-amplified-all-fragments-have-been-amplified-31-aug-04ul-dpn1-added-to-all-samples-gel-extraction-qiagen-kit-protocol-plasmid-conc-345678910-around-10ng-ul-111213141516-around-30-40-ng-ul-5-2ng-ul-7Yt-sub-8"
>
:
<br/>
9, 10, 15: Strong bands
<br/>
7, 11: weak bands
<br/>
<br/>
</span>
<span
class=
"aug-colony-pcr-attempt-1-template-solution-was-made-by-picking-colonies-and-dispersing-in-aq-medium-21-aug-gel-check-all-pcr-products-ran-on-agarose-gel-6-8-13-16-amplified-23-aug-colony-pcr-attempt-2-unamplified-sequences-reduced-annealing-temperature-to-54-53-gel-check-9-10-15-strong-bands-7-11-weak-bands-24-aug-pcr-attempt-3-all-samples-1ul-pure-plasmid-taken-as-template-25-aug-gel-check-4-5-7-8-9-14-not-amplified-27-aug-pcr-attempt-4-unamplified-samples-reduced-annealing-temperature-to-53-52-gel-check-only-9-amplified-28-aug-pcr-attempt-5-unamplified-samples-touchdown-pcr-56-52c-gel-check-4-5-unamplified-30-aug-pcr-attempt-6-unamplified-samples-50ul-reaction-separated-into-2-tubes-of-25ul-gel-check-both-fragments-4-5-amplified-all-fragments-have-been-amplified-31-aug-04ul-dpn1-added-to-all-samples-gel-extraction-qiagen-kit-protocol-plasmid-conc-345678910-around-10ng-ul-111213141516-around-30-40-ng-ul-5-2ng-ul-7Yt-sub-9"
>
24 Aug
<br/>
</span>
<span
class=
"aug-colony-pcr-attempt-1-template-solution-was-made-by-picking-colonies-and-dispersing-in-aq-medium-21-aug-gel-check-all-pcr-products-ran-on-agarose-gel-6-8-13-16-amplified-23-aug-colony-pcr-attempt-2-unamplified-sequences-reduced-annealing-temperature-to-54-53-gel-check-9-10-15-strong-bands-7-11-weak-bands-24-aug-pcr-attempt-3-all-samples-1ul-pure-plasmid-taken-as-template-25-aug-gel-check-4-5-7-8-9-14-not-amplified-27-aug-pcr-attempt-4-unamplified-samples-reduced-annealing-temperature-to-53-52-gel-check-only-9-amplified-28-aug-pcr-attempt-5-unamplified-samples-touchdown-pcr-56-52c-gel-check-4-5-unamplified-30-aug-pcr-attempt-6-unamplified-samples-50ul-reaction-separated-into-2-tubes-of-25ul-gel-check-both-fragments-4-5-amplified-all-fragments-have-been-amplified-31-aug-04ul-dpn1-added-to-all-samples-gel-extraction-qiagen-kit-protocol-plasmid-conc-345678910-around-10ng-ul-111213141516-around-30-40-ng-ul-5-2ng-ul-7Yt-sub-10"
></span>
<span
class=
"aug-colony-pcr-attempt-1-template-solution-was-made-by-picking-colonies-and-dispersing-in-aq-medium-21-aug-gel-check-all-pcr-products-ran-on-agarose-gel-6-8-13-16-amplified-23-aug-colony-pcr-attempt-2-unamplified-sequences-reduced-annealing-temperature-to-54-53-gel-check-9-10-15-strong-bands-7-11-weak-bands-24-aug-pcr-attempt-3-all-samples-1ul-pure-plasmid-taken-as-template-25-aug-gel-check-4-5-7-8-9-14-not-amplified-27-aug-pcr-attempt-4-unamplified-samples-reduced-annealing-temperature-to-53-52-gel-check-only-9-amplified-28-aug-pcr-attempt-5-unamplified-samples-touchdown-pcr-56-52c-gel-check-4-5-unamplified-30-aug-pcr-attempt-6-unamplified-samples-50ul-reaction-separated-into-2-tubes-of-25ul-gel-check-both-fragments-4-5-amplified-all-fragments-have-been-amplified-31-aug-04ul-dpn1-added-to-all-samples-gel-extraction-qiagen-kit-protocol-plasmid-conc-345678910-around-10ng-ul-111213141516-around-30-40-ng-ul-5-2ng-ul-7Yt-sub-11"
>
PCR Attempt 3
</span>
<span
class=
"aug-colony-pcr-attempt-1-template-solution-was-made-by-picking-colonies-and-dispersing-in-aq-medium-21-aug-gel-check-all-pcr-products-ran-on-agarose-gel-6-8-13-16-amplified-23-aug-colony-pcr-attempt-2-unamplified-sequences-reduced-annealing-temperature-to-54-53-gel-check-9-10-15-strong-bands-7-11-weak-bands-24-aug-pcr-attempt-3-all-samples-1ul-pure-plasmid-taken-as-template-25-aug-gel-check-4-5-7-8-9-14-not-amplified-27-aug-pcr-attempt-4-unamplified-samples-reduced-annealing-temperature-to-53-52-gel-check-only-9-amplified-28-aug-pcr-attempt-5-unamplified-samples-touchdown-pcr-56-52c-gel-check-4-5-unamplified-30-aug-pcr-attempt-6-unamplified-samples-50ul-reaction-separated-into-2-tubes-of-25ul-gel-check-both-fragments-4-5-amplified-all-fragments-have-been-amplified-31-aug-04ul-dpn1-added-to-all-samples-gel-extraction-qiagen-kit-protocol-plasmid-conc-345678910-around-10ng-ul-111213141516-around-30-40-ng-ul-5-2ng-ul-7Yt-sub-12"
>
(All Samples) : 1uL pure plasmid taken as template
<br/>
</span>
<span
class=
"aug-colony-pcr-attempt-1-template-solution-was-made-by-picking-colonies-and-dispersing-in-aq-medium-21-aug-gel-check-all-pcr-products-ran-on-agarose-gel-6-8-13-16-amplified-23-aug-colony-pcr-attempt-2-unamplified-sequences-reduced-annealing-temperature-to-54-53-gel-check-9-10-15-strong-bands-7-11-weak-bands-24-aug-pcr-attempt-3-all-samples-1ul-pure-plasmid-taken-as-template-25-aug-gel-check-4-5-7-8-9-14-not-amplified-27-aug-pcr-attempt-4-unamplified-samples-reduced-annealing-temperature-to-53-52-gel-check-only-9-amplified-28-aug-pcr-attempt-5-unamplified-samples-touchdown-pcr-56-52c-gel-check-4-5-unamplified-30-aug-pcr-attempt-6-unamplified-samples-50ul-reaction-separated-into-2-tubes-of-25ul-gel-check-both-fragments-4-5-amplified-all-fragments-have-been-amplified-31-aug-04ul-dpn1-added-to-all-samples-gel-extraction-qiagen-kit-protocol-plasmid-conc-345678910-around-10ng-ul-111213141516-around-30-40-ng-ul-5-2ng-ul-7Yt-sub-13"
>
<br/>
25 Aug
<br/>
Gel Check
</span>
<span
class=
"aug-colony-pcr-attempt-1-template-solution-was-made-by-picking-colonies-and-dispersing-in-aq-medium-21-aug-gel-check-all-pcr-products-ran-on-agarose-gel-6-8-13-16-amplified-23-aug-colony-pcr-attempt-2-unamplified-sequences-reduced-annealing-temperature-to-54-53-gel-check-9-10-15-strong-bands-7-11-weak-bands-24-aug-pcr-attempt-3-all-samples-1ul-pure-plasmid-taken-as-template-25-aug-gel-check-4-5-7-8-9-14-not-amplified-27-aug-pcr-attempt-4-unamplified-samples-reduced-annealing-temperature-to-53-52-gel-check-only-9-amplified-28-aug-pcr-attempt-5-unamplified-samples-touchdown-pcr-56-52c-gel-check-4-5-unamplified-30-aug-pcr-attempt-6-unamplified-samples-50ul-reaction-separated-into-2-tubes-of-25ul-gel-check-both-fragments-4-5-amplified-all-fragments-have-been-amplified-31-aug-04ul-dpn1-added-to-all-samples-gel-extraction-qiagen-kit-protocol-plasmid-conc-345678910-around-10ng-ul-111213141516-around-30-40-ng-ul-5-2ng-ul-7Yt-sub-14"
>
: 4, 5, 7, 8, 9 14 Not amplified
<br/>
<br/>
</span>
<span
class=
"aug-colony-pcr-attempt-1-template-solution-was-made-by-picking-colonies-and-dispersing-in-aq-medium-21-aug-gel-check-all-pcr-products-ran-on-agarose-gel-6-8-13-16-amplified-23-aug-colony-pcr-attempt-2-unamplified-sequences-reduced-annealing-temperature-to-54-53-gel-check-9-10-15-strong-bands-7-11-weak-bands-24-aug-pcr-attempt-3-all-samples-1ul-pure-plasmid-taken-as-template-25-aug-gel-check-4-5-7-8-9-14-not-amplified-27-aug-pcr-attempt-4-unamplified-samples-reduced-annealing-temperature-to-53-52-gel-check-only-9-amplified-28-aug-pcr-attempt-5-unamplified-samples-touchdown-pcr-56-52c-gel-check-4-5-unamplified-30-aug-pcr-attempt-6-unamplified-samples-50ul-reaction-separated-into-2-tubes-of-25ul-gel-check-both-fragments-4-5-amplified-all-fragments-have-been-amplified-31-aug-04ul-dpn1-added-to-all-samples-gel-extraction-qiagen-kit-protocol-plasmid-conc-345678910-around-10ng-ul-111213141516-around-30-40-ng-ul-5-2ng-ul-7Yt-sub-15"
>
27 Aug
<br/>
PCR Attempt 4
</span>
<span
class=
"aug-colony-pcr-attempt-1-template-solution-was-made-by-picking-colonies-and-dispersing-in-aq-medium-21-aug-gel-check-all-pcr-products-ran-on-agarose-gel-6-8-13-16-amplified-23-aug-colony-pcr-attempt-2-unamplified-sequences-reduced-annealing-temperature-to-54-53-gel-check-9-10-15-strong-bands-7-11-weak-bands-24-aug-pcr-attempt-3-all-samples-1ul-pure-plasmid-taken-as-template-25-aug-gel-check-4-5-7-8-9-14-not-amplified-27-aug-pcr-attempt-4-unamplified-samples-reduced-annealing-temperature-to-53-52-gel-check-only-9-amplified-28-aug-pcr-attempt-5-unamplified-samples-touchdown-pcr-56-52c-gel-check-4-5-unamplified-30-aug-pcr-attempt-6-unamplified-samples-50ul-reaction-separated-into-2-tubes-of-25ul-gel-check-both-fragments-4-5-amplified-all-fragments-have-been-amplified-31-aug-04ul-dpn1-added-to-all-samples-gel-extraction-qiagen-kit-protocol-plasmid-conc-345678910-around-10ng-ul-111213141516-around-30-40-ng-ul-5-2ng-ul-7Yt-sub-16"
>
(Unamplified samples) : Reduced annealing temperature to 53, 52
<br/>
</span>
<span
class=
"aug-colony-pcr-attempt-1-template-solution-was-made-by-picking-colonies-and-dispersing-in-aq-medium-21-aug-gel-check-all-pcr-products-ran-on-agarose-gel-6-8-13-16-amplified-23-aug-colony-pcr-attempt-2-unamplified-sequences-reduced-annealing-temperature-to-54-53-gel-check-9-10-15-strong-bands-7-11-weak-bands-24-aug-pcr-attempt-3-all-samples-1ul-pure-plasmid-taken-as-template-25-aug-gel-check-4-5-7-8-9-14-not-amplified-27-aug-pcr-attempt-4-unamplified-samples-reduced-annealing-temperature-to-53-52-gel-check-only-9-amplified-28-aug-pcr-attempt-5-unamplified-samples-touchdown-pcr-56-52c-gel-check-4-5-unamplified-30-aug-pcr-attempt-6-unamplified-samples-50ul-reaction-separated-into-2-tubes-of-25ul-gel-check-both-fragments-4-5-amplified-all-fragments-have-been-amplified-31-aug-04ul-dpn1-added-to-all-samples-gel-extraction-qiagen-kit-protocol-plasmid-conc-345678910-around-10ng-ul-111213141516-around-30-40-ng-ul-5-2ng-ul-7Yt-sub-17"
>
Gel Check
</span>
<span
class=
"aug-colony-pcr-attempt-1-template-solution-was-made-by-picking-colonies-and-dispersing-in-aq-medium-21-aug-gel-check-all-pcr-products-ran-on-agarose-gel-6-8-13-16-amplified-23-aug-colony-pcr-attempt-2-unamplified-sequences-reduced-annealing-temperature-to-54-53-gel-check-9-10-15-strong-bands-7-11-weak-bands-24-aug-pcr-attempt-3-all-samples-1ul-pure-plasmid-taken-as-template-25-aug-gel-check-4-5-7-8-9-14-not-amplified-27-aug-pcr-attempt-4-unamplified-samples-reduced-annealing-temperature-to-53-52-gel-check-only-9-amplified-28-aug-pcr-attempt-5-unamplified-samples-touchdown-pcr-56-52c-gel-check-4-5-unamplified-30-aug-pcr-attempt-6-unamplified-samples-50ul-reaction-separated-into-2-tubes-of-25ul-gel-check-both-fragments-4-5-amplified-all-fragments-have-been-amplified-31-aug-04ul-dpn1-added-to-all-samples-gel-extraction-qiagen-kit-protocol-plasmid-conc-345678910-around-10ng-ul-111213141516-around-30-40-ng-ul-5-2ng-ul-7Yt-sub-18"
>
: only 9 Amplified
<br/>
<br/>
</span>
<span
class=
"aug-colony-pcr-attempt-1-template-solution-was-made-by-picking-colonies-and-dispersing-in-aq-medium-21-aug-gel-check-all-pcr-products-ran-on-agarose-gel-6-8-13-16-amplified-23-aug-colony-pcr-attempt-2-unamplified-sequences-reduced-annealing-temperature-to-54-53-gel-check-9-10-15-strong-bands-7-11-weak-bands-24-aug-pcr-attempt-3-all-samples-1ul-pure-plasmid-taken-as-template-25-aug-gel-check-4-5-7-8-9-14-not-amplified-27-aug-pcr-attempt-4-unamplified-samples-reduced-annealing-temperature-to-53-52-gel-check-only-9-amplified-28-aug-pcr-attempt-5-unamplified-samples-touchdown-pcr-56-52c-gel-check-4-5-unamplified-30-aug-pcr-attempt-6-unamplified-samples-50ul-reaction-separated-into-2-tubes-of-25ul-gel-check-both-fragments-4-5-amplified-all-fragments-have-been-amplified-31-aug-04ul-dpn1-added-to-all-samples-gel-extraction-qiagen-kit-protocol-plasmid-conc-345678910-around-10ng-ul-111213141516-around-30-40-ng-ul-5-2ng-ul-7Yt-sub-19"
>
28 Aug
<br/>
PCR Attempt 5
</span>
<span
class=
"aug-colony-pcr-attempt-1-template-solution-was-made-by-picking-colonies-and-dispersing-in-aq-medium-21-aug-gel-check-all-pcr-products-ran-on-agarose-gel-6-8-13-16-amplified-23-aug-colony-pcr-attempt-2-unamplified-sequences-reduced-annealing-temperature-to-54-53-gel-check-9-10-15-strong-bands-7-11-weak-bands-24-aug-pcr-attempt-3-all-samples-1ul-pure-plasmid-taken-as-template-25-aug-gel-check-4-5-7-8-9-14-not-amplified-27-aug-pcr-attempt-4-unamplified-samples-reduced-annealing-temperature-to-53-52-gel-check-only-9-amplified-28-aug-pcr-attempt-5-unamplified-samples-touchdown-pcr-56-52c-gel-check-4-5-unamplified-30-aug-pcr-attempt-6-unamplified-samples-50ul-reaction-separated-into-2-tubes-of-25ul-gel-check-both-fragments-4-5-amplified-all-fragments-have-been-amplified-31-aug-04ul-dpn1-added-to-all-samples-gel-extraction-qiagen-kit-protocol-plasmid-conc-345678910-around-10ng-ul-111213141516-around-30-40-ng-ul-5-2ng-ul-7Yt-sub-20"
>
(Unamplified samples) : Touchdown PCR 56-52C
<br/>
</span>
<span
class=
"aug-colony-pcr-attempt-1-template-solution-was-made-by-picking-colonies-and-dispersing-in-aq-medium-21-aug-gel-check-all-pcr-products-ran-on-agarose-gel-6-8-13-16-amplified-23-aug-colony-pcr-attempt-2-unamplified-sequences-reduced-annealing-temperature-to-54-53-gel-check-9-10-15-strong-bands-7-11-weak-bands-24-aug-pcr-attempt-3-all-samples-1ul-pure-plasmid-taken-as-template-25-aug-gel-check-4-5-7-8-9-14-not-amplified-27-aug-pcr-attempt-4-unamplified-samples-reduced-annealing-temperature-to-53-52-gel-check-only-9-amplified-28-aug-pcr-attempt-5-unamplified-samples-touchdown-pcr-56-52c-gel-check-4-5-unamplified-30-aug-pcr-attempt-6-unamplified-samples-50ul-reaction-separated-into-2-tubes-of-25ul-gel-check-both-fragments-4-5-amplified-all-fragments-have-been-amplified-31-aug-04ul-dpn1-added-to-all-samples-gel-extraction-qiagen-kit-protocol-plasmid-conc-345678910-around-10ng-ul-111213141516-around-30-40-ng-ul-5-2ng-ul-7Yt-sub-21"
>
Gel Check
</span>
<span
class=
"aug-colony-pcr-attempt-1-template-solution-was-made-by-picking-colonies-and-dispersing-in-aq-medium-21-aug-gel-check-all-pcr-products-ran-on-agarose-gel-6-8-13-16-amplified-23-aug-colony-pcr-attempt-2-unamplified-sequences-reduced-annealing-temperature-to-54-53-gel-check-9-10-15-strong-bands-7-11-weak-bands-24-aug-pcr-attempt-3-all-samples-1ul-pure-plasmid-taken-as-template-25-aug-gel-check-4-5-7-8-9-14-not-amplified-27-aug-pcr-attempt-4-unamplified-samples-reduced-annealing-temperature-to-53-52-gel-check-only-9-amplified-28-aug-pcr-attempt-5-unamplified-samples-touchdown-pcr-56-52c-gel-check-4-5-unamplified-30-aug-pcr-attempt-6-unamplified-samples-50ul-reaction-separated-into-2-tubes-of-25ul-gel-check-both-fragments-4-5-amplified-all-fragments-have-been-amplified-31-aug-04ul-dpn1-added-to-all-samples-gel-extraction-qiagen-kit-protocol-plasmid-conc-345678910-around-10ng-ul-111213141516-around-30-40-ng-ul-5-2ng-ul-7Yt-sub-22"
>
: 4, 5 Unamplified
<br/>
<br/>
</span>
<span
class=
"aug-colony-pcr-attempt-1-template-solution-was-made-by-picking-colonies-and-dispersing-in-aq-medium-21-aug-gel-check-all-pcr-products-ran-on-agarose-gel-6-8-13-16-amplified-23-aug-colony-pcr-attempt-2-unamplified-sequences-reduced-annealing-temperature-to-54-53-gel-check-9-10-15-strong-bands-7-11-weak-bands-24-aug-pcr-attempt-3-all-samples-1ul-pure-plasmid-taken-as-template-25-aug-gel-check-4-5-7-8-9-14-not-amplified-27-aug-pcr-attempt-4-unamplified-samples-reduced-annealing-temperature-to-53-52-gel-check-only-9-amplified-28-aug-pcr-attempt-5-unamplified-samples-touchdown-pcr-56-52c-gel-check-4-5-unamplified-30-aug-pcr-attempt-6-unamplified-samples-50ul-reaction-separated-into-2-tubes-of-25ul-gel-check-both-fragments-4-5-amplified-all-fragments-have-been-amplified-31-aug-04ul-dpn1-added-to-all-samples-gel-extraction-qiagen-kit-protocol-plasmid-conc-345678910-around-10ng-ul-111213141516-around-30-40-ng-ul-5-2ng-ul-7Yt-sub-23"
>
30 Aug
<br/>
PCR Attempt 6
</span>
<span
class=
"aug-colony-pcr-attempt-1-template-solution-was-made-by-picking-colonies-and-dispersing-in-aq-medium-21-aug-gel-check-all-pcr-products-ran-on-agarose-gel-6-8-13-16-amplified-23-aug-colony-pcr-attempt-2-unamplified-sequences-reduced-annealing-temperature-to-54-53-gel-check-9-10-15-strong-bands-7-11-weak-bands-24-aug-pcr-attempt-3-all-samples-1ul-pure-plasmid-taken-as-template-25-aug-gel-check-4-5-7-8-9-14-not-amplified-27-aug-pcr-attempt-4-unamplified-samples-reduced-annealing-temperature-to-53-52-gel-check-only-9-amplified-28-aug-pcr-attempt-5-unamplified-samples-touchdown-pcr-56-52c-gel-check-4-5-unamplified-30-aug-pcr-attempt-6-unamplified-samples-50ul-reaction-separated-into-2-tubes-of-25ul-gel-check-both-fragments-4-5-amplified-all-fragments-have-been-amplified-31-aug-04ul-dpn1-added-to-all-samples-gel-extraction-qiagen-kit-protocol-plasmid-conc-345678910-around-10ng-ul-111213141516-around-30-40-ng-ul-5-2ng-ul-7Yt-sub-24"
>
(Unamplified samples) : 50uL reaction separated into 2 tubes of 25uL
<br/>
</span>
<span
class=
"aug-colony-pcr-attempt-1-template-solution-was-made-by-picking-colonies-and-dispersing-in-aq-medium-21-aug-gel-check-all-pcr-products-ran-on-agarose-gel-6-8-13-16-amplified-23-aug-colony-pcr-attempt-2-unamplified-sequences-reduced-annealing-temperature-to-54-53-gel-check-9-10-15-strong-bands-7-11-weak-bands-24-aug-pcr-attempt-3-all-samples-1ul-pure-plasmid-taken-as-template-25-aug-gel-check-4-5-7-8-9-14-not-amplified-27-aug-pcr-attempt-4-unamplified-samples-reduced-annealing-temperature-to-53-52-gel-check-only-9-amplified-28-aug-pcr-attempt-5-unamplified-samples-touchdown-pcr-56-52c-gel-check-4-5-unamplified-30-aug-pcr-attempt-6-unamplified-samples-50ul-reaction-separated-into-2-tubes-of-25ul-gel-check-both-fragments-4-5-amplified-all-fragments-have-been-amplified-31-aug-04ul-dpn1-added-to-all-samples-gel-extraction-qiagen-kit-protocol-plasmid-conc-345678910-around-10ng-ul-111213141516-around-30-40-ng-ul-5-2ng-ul-7Yt-sub-25"
>
Gel Check
</span>
<span
class=
"aug-colony-pcr-attempt-1-template-solution-was-made-by-picking-colonies-and-dispersing-in-aq-medium-21-aug-gel-check-all-pcr-products-ran-on-agarose-gel-6-8-13-16-amplified-23-aug-colony-pcr-attempt-2-unamplified-sequences-reduced-annealing-temperature-to-54-53-gel-check-9-10-15-strong-bands-7-11-weak-bands-24-aug-pcr-attempt-3-all-samples-1ul-pure-plasmid-taken-as-template-25-aug-gel-check-4-5-7-8-9-14-not-amplified-27-aug-pcr-attempt-4-unamplified-samples-reduced-annealing-temperature-to-53-52-gel-check-only-9-amplified-28-aug-pcr-attempt-5-unamplified-samples-touchdown-pcr-56-52c-gel-check-4-5-unamplified-30-aug-pcr-attempt-6-unamplified-samples-50ul-reaction-separated-into-2-tubes-of-25ul-gel-check-both-fragments-4-5-amplified-all-fragments-have-been-amplified-31-aug-04ul-dpn1-added-to-all-samples-gel-extraction-qiagen-kit-protocol-plasmid-conc-345678910-around-10ng-ul-111213141516-around-30-40-ng-ul-5-2ng-ul-7Yt-sub-26"
>
: both fragments (4, 5) amplified
<br/>
<br/>
</span>
<span
class=
"aug-colony-pcr-attempt-1-template-solution-was-made-by-picking-colonies-and-dispersing-in-aq-medium-21-aug-gel-check-all-pcr-products-ran-on-agarose-gel-6-8-13-16-amplified-23-aug-colony-pcr-attempt-2-unamplified-sequences-reduced-annealing-temperature-to-54-53-gel-check-9-10-15-strong-bands-7-11-weak-bands-24-aug-pcr-attempt-3-all-samples-1ul-pure-plasmid-taken-as-template-25-aug-gel-check-4-5-7-8-9-14-not-amplified-27-aug-pcr-attempt-4-unamplified-samples-reduced-annealing-temperature-to-53-52-gel-check-only-9-amplified-28-aug-pcr-attempt-5-unamplified-samples-touchdown-pcr-56-52c-gel-check-4-5-unamplified-30-aug-pcr-attempt-6-unamplified-samples-50ul-reaction-separated-into-2-tubes-of-25ul-gel-check-both-fragments-4-5-amplified-all-fragments-have-been-amplified-31-aug-04ul-dpn1-added-to-all-samples-gel-extraction-qiagen-kit-protocol-plasmid-conc-345678910-around-10ng-ul-111213141516-around-30-40-ng-ul-5-2ng-ul-7Yt-sub-27"
>
All fragments have been amplified!
<br/>
</span>
<span
class=
"aug-colony-pcr-attempt-1-template-solution-was-made-by-picking-colonies-and-dispersing-in-aq-medium-21-aug-gel-check-all-pcr-products-ran-on-agarose-gel-6-8-13-16-amplified-23-aug-colony-pcr-attempt-2-unamplified-sequences-reduced-annealing-temperature-to-54-53-gel-check-9-10-15-strong-bands-7-11-weak-bands-24-aug-pcr-attempt-3-all-samples-1ul-pure-plasmid-taken-as-template-25-aug-gel-check-4-5-7-8-9-14-not-amplified-27-aug-pcr-attempt-4-unamplified-samples-reduced-annealing-temperature-to-53-52-gel-check-only-9-amplified-28-aug-pcr-attempt-5-unamplified-samples-touchdown-pcr-56-52c-gel-check-4-5-unamplified-30-aug-pcr-attempt-6-unamplified-samples-50ul-reaction-separated-into-2-tubes-of-25ul-gel-check-both-fragments-4-5-amplified-all-fragments-have-been-amplified-31-aug-04ul-dpn1-added-to-all-samples-gel-extraction-qiagen-kit-protocol-plasmid-conc-345678910-around-10ng-ul-111213141516-around-30-40-ng-ul-5-2ng-ul-7Yt-sub-28"
>
<br/>
</span>
<span
class=
"aug-colony-pcr-attempt-1-template-solution-was-made-by-picking-colonies-and-dispersing-in-aq-medium-21-aug-gel-check-all-pcr-products-ran-on-agarose-gel-6-8-13-16-amplified-23-aug-colony-pcr-attempt-2-unamplified-sequences-reduced-annealing-temperature-to-54-53-gel-check-9-10-15-strong-bands-7-11-weak-bands-24-aug-pcr-attempt-3-all-samples-1ul-pure-plasmid-taken-as-template-25-aug-gel-check-4-5-7-8-9-14-not-amplified-27-aug-pcr-attempt-4-unamplified-samples-reduced-annealing-temperature-to-53-52-gel-check-only-9-amplified-28-aug-pcr-attempt-5-unamplified-samples-touchdown-pcr-56-52c-gel-check-4-5-unamplified-30-aug-pcr-attempt-6-unamplified-samples-50ul-reaction-separated-into-2-tubes-of-25ul-gel-check-both-fragments-4-5-amplified-all-fragments-have-been-amplified-31-aug-04ul-dpn1-added-to-all-samples-gel-extraction-qiagen-kit-protocol-plasmid-conc-345678910-around-10ng-ul-111213141516-around-30-40-ng-ul-5-2ng-ul-7Yt-sub-29"
>
31 Aug
<br/>
</span>
<span
class=
"aug-colony-pcr-attempt-1-template-solution-was-made-by-picking-colonies-and-dispersing-in-aq-medium-21-aug-gel-check-all-pcr-products-ran-on-agarose-gel-6-8-13-16-amplified-23-aug-colony-pcr-attempt-2-unamplified-sequences-reduced-annealing-temperature-to-54-53-gel-check-9-10-15-strong-bands-7-11-weak-bands-24-aug-pcr-attempt-3-all-samples-1ul-pure-plasmid-taken-as-template-25-aug-gel-check-4-5-7-8-9-14-not-amplified-27-aug-pcr-attempt-4-unamplified-samples-reduced-annealing-temperature-to-53-52-gel-check-only-9-amplified-28-aug-pcr-attempt-5-unamplified-samples-touchdown-pcr-56-52c-gel-check-4-5-unamplified-30-aug-pcr-attempt-6-unamplified-samples-50ul-reaction-separated-into-2-tubes-of-25ul-gel-check-both-fragments-4-5-amplified-all-fragments-have-been-amplified-31-aug-04ul-dpn1-added-to-all-samples-gel-extraction-qiagen-kit-protocol-plasmid-conc-345678910-around-10ng-ul-111213141516-around-30-40-ng-ul-5-2ng-ul-7Yt-sub-30"
>
0.4uL Dpn1 added to all samples
<br/>
</span>
<span
class=
"aug-colony-pcr-attempt-1-template-solution-was-made-by-picking-colonies-and-dispersing-in-aq-medium-21-aug-gel-check-all-pcr-products-ran-on-agarose-gel-6-8-13-16-amplified-23-aug-colony-pcr-attempt-2-unamplified-sequences-reduced-annealing-temperature-to-54-53-gel-check-9-10-15-strong-bands-7-11-weak-bands-24-aug-pcr-attempt-3-all-samples-1ul-pure-plasmid-taken-as-template-25-aug-gel-check-4-5-7-8-9-14-not-amplified-27-aug-pcr-attempt-4-unamplified-samples-reduced-annealing-temperature-to-53-52-gel-check-only-9-amplified-28-aug-pcr-attempt-5-unamplified-samples-touchdown-pcr-56-52c-gel-check-4-5-unamplified-30-aug-pcr-attempt-6-unamplified-samples-50ul-reaction-separated-into-2-tubes-of-25ul-gel-check-both-fragments-4-5-amplified-all-fragments-have-been-amplified-31-aug-04ul-dpn1-added-to-all-samples-gel-extraction-qiagen-kit-protocol-plasmid-conc-345678910-around-10ng-ul-111213141516-around-30-40-ng-ul-5-2ng-ul-7Yt-sub-31"
>
Gel Extraction
</span>
<span
class=
"aug-colony-pcr-attempt-1-template-solution-was-made-by-picking-colonies-and-dispersing-in-aq-medium-21-aug-gel-check-all-pcr-products-ran-on-agarose-gel-6-8-13-16-amplified-23-aug-colony-pcr-attempt-2-unamplified-sequences-reduced-annealing-temperature-to-54-53-gel-check-9-10-15-strong-bands-7-11-weak-bands-24-aug-pcr-attempt-3-all-samples-1ul-pure-plasmid-taken-as-template-25-aug-gel-check-4-5-7-8-9-14-not-amplified-27-aug-pcr-attempt-4-unamplified-samples-reduced-annealing-temperature-to-53-52-gel-check-only-9-amplified-28-aug-pcr-attempt-5-unamplified-samples-touchdown-pcr-56-52c-gel-check-4-5-unamplified-30-aug-pcr-attempt-6-unamplified-samples-50ul-reaction-separated-into-2-tubes-of-25ul-gel-check-both-fragments-4-5-amplified-all-fragments-have-been-amplified-31-aug-04ul-dpn1-added-to-all-samples-gel-extraction-qiagen-kit-protocol-plasmid-conc-345678910-around-10ng-ul-111213141516-around-30-40-ng-ul-5-2ng-ul-7Yt-sub-32"
>
: (Qiagen Kit protocol)
<br/>
Plasmid conc :
<br/>
3,4,5,6,7,8,9,10 - around 10ng/uL.
<br/>
11,12,13,14,15,16 - around 30-40 ng/uL
<br/>
5 - 2ng/ul
</span>
</p>
<p
class=
"sequences-are-referred-to-by-serial-numbers-optimization-algorithms-used-to-generate-the-sequence-sa-simulated-annealing-gd-gradient-descent-ga-genetic-algorithm-h-l-high-low-expressing-ZDA"
id=
"539:54"
>
Sequences are referred to by serial numbers
<br/>
<br/>
Optimization Algorithms used to generate the sequence:
<br/>
SA - Simulated Annealing
<br/>
GD - Gradient Descent
<br/>
GA - Genetic Algorithm
<br/>
<br/>
H, L = High, Low Expressing
</p>
</div>
</div>
<div
class=
"group-43-jmr"
id=
"400:46"
>
<p
class=
"september-ffW"
id=
"400:48"
>
September
</p>
<p
class=
"sept-transformation-plating-electroporation-was-done-for-all-the-samples-in-l-lactis-competent-cells-samples-were-incubated-for-1hr-after-which-they-were-spread-plated-and-incubated-for-24-hours-3-sept-colonies-in-all-plates-except-samples-4-and-14-10-colonies-were-observed-in-plate-3-colonies-were-picked-and-cultured-overnight-2-cultures-from-each-non-randomized-sample-all-10-cultures-of-each-colony-from-the-randomized-sample-3-4-sept-glycerol-added-and-strains-stored-in-80c-5-sept-transformation-and-plating-attempt-2-samples-3414-working-to-obtain-more-strains-of-randomized-sample-3-6-sept-no-colonies-observed-8-sept-transformation-and-plating-attempt-4-samples-3414-9-sept-no-colonies-observed-11-sept-transformation-and-plating-attempt-5-samples-3414-slightly-reduced-antibiotic-concentration-12-sept-golden-gate-assembly-set-up-from-left-over-plasmid-for-3414-14-sept-transformation-and-plating-attempt-6-samples-3414-15-sept-1-colony-for-sample-3-none-for-4-and-14-17-sept-transformation-and-plating-attempt-7-samples-3414-18-sept-2-colonies-in-sample-3-none-for-4-and-14-21-sept-all-strains-13-variants-from-randomized-sample-3-11-model-generated-sequences-streaked-on-agar-plates-22-sept-colonies-picked-and-overnight-1ml-cultures-made-and-incubated-for-14-hrs-23-sept-fluorescence-assay-first-batch-of-results-obtained-25-sept-colonies-picked-from-streaked-plates-and-cultured-overnight-26-sept-plasmid-isolation-qiagen-kit-protocol-plasmid-concentration-obtained-were-very-poor-b3N"
id=
"400:49"
>
<span
class=
"sept-transformation-plating-electroporation-was-done-for-all-the-samples-in-l-lactis-competent-cells-samples-were-incubated-for-1hr-after-which-they-were-spread-plated-and-incubated-for-24-hours-3-sept-colonies-in-all-plates-except-samples-4-and-14-10-colonies-were-observed-in-plate-3-colonies-were-picked-and-cultured-overnight-2-cultures-from-each-non-randomized-sample-all-10-cultures-of-each-colony-from-the-randomized-sample-3-4-sept-glycerol-added-and-strains-stored-in-80c-5-sept-transformation-and-plating-attempt-2-samples-3414-working-to-obtain-more-strains-of-randomized-sample-3-6-sept-no-colonies-observed-8-sept-transformation-and-plating-attempt-4-samples-3414-9-sept-no-colonies-observed-11-sept-transformation-and-plating-attempt-5-samples-3414-slightly-reduced-antibiotic-concentration-12-sept-golden-gate-assembly-set-up-from-left-over-plasmid-for-3414-14-sept-transformation-and-plating-attempt-6-samples-3414-15-sept-1-colony-for-sample-3-none-for-4-and-14-17-sept-transformation-and-plating-attempt-7-samples-3414-18-sept-2-colonies-in-sample-3-none-for-4-and-14-21-sept-all-strains-13-variants-from-randomized-sample-3-11-model-generated-sequences-streaked-on-agar-plates-22-sept-colonies-picked-and-overnight-1ml-cultures-made-and-incubated-for-14-hrs-23-sept-fluorescence-assay-first-batch-of-results-obtained-25-sept-colonies-picked-from-streaked-plates-and-cultured-overnight-26-sept-plasmid-isolation-qiagen-kit-protocol-plasmid-concentration-obtained-were-very-poor-b3N-sub-0"
>
2 Sept
<br/>
</span>
<span
class=
"sept-transformation-plating-electroporation-was-done-for-all-the-samples-in-l-lactis-competent-cells-samples-were-incubated-for-1hr-after-which-they-were-spread-plated-and-incubated-for-24-hours-3-sept-colonies-in-all-plates-except-samples-4-and-14-10-colonies-were-observed-in-plate-3-colonies-were-picked-and-cultured-overnight-2-cultures-from-each-non-randomized-sample-all-10-cultures-of-each-colony-from-the-randomized-sample-3-4-sept-glycerol-added-and-strains-stored-in-80c-5-sept-transformation-and-plating-attempt-2-samples-3414-working-to-obtain-more-strains-of-randomized-sample-3-6-sept-no-colonies-observed-8-sept-transformation-and-plating-attempt-4-samples-3414-9-sept-no-colonies-observed-11-sept-transformation-and-plating-attempt-5-samples-3414-slightly-reduced-antibiotic-concentration-12-sept-golden-gate-assembly-set-up-from-left-over-plasmid-for-3414-14-sept-transformation-and-plating-attempt-6-samples-3414-15-sept-1-colony-for-sample-3-none-for-4-and-14-17-sept-transformation-and-plating-attempt-7-samples-3414-18-sept-2-colonies-in-sample-3-none-for-4-and-14-21-sept-all-strains-13-variants-from-randomized-sample-3-11-model-generated-sequences-streaked-on-agar-plates-22-sept-colonies-picked-and-overnight-1ml-cultures-made-and-incubated-for-14-hrs-23-sept-fluorescence-assay-first-batch-of-results-obtained-25-sept-colonies-picked-from-streaked-plates-and-cultured-overnight-26-sept-plasmid-isolation-qiagen-kit-protocol-plasmid-concentration-obtained-were-very-poor-b3N-sub-1"
>
Transformation + Plating
<br/>
Electroporation was done for all the samples in
</span>
<span
class=
"sept-transformation-plating-electroporation-was-done-for-all-the-samples-in-l-lactis-competent-cells-samples-were-incubated-for-1hr-after-which-they-were-spread-plated-and-incubated-for-24-hours-3-sept-colonies-in-all-plates-except-samples-4-and-14-10-colonies-were-observed-in-plate-3-colonies-were-picked-and-cultured-overnight-2-cultures-from-each-non-randomized-sample-all-10-cultures-of-each-colony-from-the-randomized-sample-3-4-sept-glycerol-added-and-strains-stored-in-80c-5-sept-transformation-and-plating-attempt-2-samples-3414-working-to-obtain-more-strains-of-randomized-sample-3-6-sept-no-colonies-observed-8-sept-transformation-and-plating-attempt-4-samples-3414-9-sept-no-colonies-observed-11-sept-transformation-and-plating-attempt-5-samples-3414-slightly-reduced-antibiotic-concentration-12-sept-golden-gate-assembly-set-up-from-left-over-plasmid-for-3414-14-sept-transformation-and-plating-attempt-6-samples-3414-15-sept-1-colony-for-sample-3-none-for-4-and-14-17-sept-transformation-and-plating-attempt-7-samples-3414-18-sept-2-colonies-in-sample-3-none-for-4-and-14-21-sept-all-strains-13-variants-from-randomized-sample-3-11-model-generated-sequences-streaked-on-agar-plates-22-sept-colonies-picked-and-overnight-1ml-cultures-made-and-incubated-for-14-hrs-23-sept-fluorescence-assay-first-batch-of-results-obtained-25-sept-colonies-picked-from-streaked-plates-and-cultured-overnight-26-sept-plasmid-isolation-qiagen-kit-protocol-plasmid-concentration-obtained-were-very-poor-b3N-sub-2"
>
L. lactis
</span>
<span
class=
"sept-transformation-plating-electroporation-was-done-for-all-the-samples-in-l-lactis-competent-cells-samples-were-incubated-for-1hr-after-which-they-were-spread-plated-and-incubated-for-24-hours-3-sept-colonies-in-all-plates-except-samples-4-and-14-10-colonies-were-observed-in-plate-3-colonies-were-picked-and-cultured-overnight-2-cultures-from-each-non-randomized-sample-all-10-cultures-of-each-colony-from-the-randomized-sample-3-4-sept-glycerol-added-and-strains-stored-in-80c-5-sept-transformation-and-plating-attempt-2-samples-3414-working-to-obtain-more-strains-of-randomized-sample-3-6-sept-no-colonies-observed-8-sept-transformation-and-plating-attempt-4-samples-3414-9-sept-no-colonies-observed-11-sept-transformation-and-plating-attempt-5-samples-3414-slightly-reduced-antibiotic-concentration-12-sept-golden-gate-assembly-set-up-from-left-over-plasmid-for-3414-14-sept-transformation-and-plating-attempt-6-samples-3414-15-sept-1-colony-for-sample-3-none-for-4-and-14-17-sept-transformation-and-plating-attempt-7-samples-3414-18-sept-2-colonies-in-sample-3-none-for-4-and-14-21-sept-all-strains-13-variants-from-randomized-sample-3-11-model-generated-sequences-streaked-on-agar-plates-22-sept-colonies-picked-and-overnight-1ml-cultures-made-and-incubated-for-14-hrs-23-sept-fluorescence-assay-first-batch-of-results-obtained-25-sept-colonies-picked-from-streaked-plates-and-cultured-overnight-26-sept-plasmid-isolation-qiagen-kit-protocol-plasmid-concentration-obtained-were-very-poor-b3N-sub-3"
>
competent cells.
<br/>
Samples were incubated for 1hr after which they were spread plated and incubated for 24 hours.
<br/>
<br/>
</span>
<span
class=
"sept-transformation-plating-electroporation-was-done-for-all-the-samples-in-l-lactis-competent-cells-samples-were-incubated-for-1hr-after-which-they-were-spread-plated-and-incubated-for-24-hours-3-sept-colonies-in-all-plates-except-samples-4-and-14-10-colonies-were-observed-in-plate-3-colonies-were-picked-and-cultured-overnight-2-cultures-from-each-non-randomized-sample-all-10-cultures-of-each-colony-from-the-randomized-sample-3-4-sept-glycerol-added-and-strains-stored-in-80c-5-sept-transformation-and-plating-attempt-2-samples-3414-working-to-obtain-more-strains-of-randomized-sample-3-6-sept-no-colonies-observed-8-sept-transformation-and-plating-attempt-4-samples-3414-9-sept-no-colonies-observed-11-sept-transformation-and-plating-attempt-5-samples-3414-slightly-reduced-antibiotic-concentration-12-sept-golden-gate-assembly-set-up-from-left-over-plasmid-for-3414-14-sept-transformation-and-plating-attempt-6-samples-3414-15-sept-1-colony-for-sample-3-none-for-4-and-14-17-sept-transformation-and-plating-attempt-7-samples-3414-18-sept-2-colonies-in-sample-3-none-for-4-and-14-21-sept-all-strains-13-variants-from-randomized-sample-3-11-model-generated-sequences-streaked-on-agar-plates-22-sept-colonies-picked-and-overnight-1ml-cultures-made-and-incubated-for-14-hrs-23-sept-fluorescence-assay-first-batch-of-results-obtained-25-sept-colonies-picked-from-streaked-plates-and-cultured-overnight-26-sept-plasmid-isolation-qiagen-kit-protocol-plasmid-concentration-obtained-were-very-poor-b3N-sub-4"
>
3 Sept
<br/>
</span>
<span
class=
"sept-transformation-plating-electroporation-was-done-for-all-the-samples-in-l-lactis-competent-cells-samples-were-incubated-for-1hr-after-which-they-were-spread-plated-and-incubated-for-24-hours-3-sept-colonies-in-all-plates-except-samples-4-and-14-10-colonies-were-observed-in-plate-3-colonies-were-picked-and-cultured-overnight-2-cultures-from-each-non-randomized-sample-all-10-cultures-of-each-colony-from-the-randomized-sample-3-4-sept-glycerol-added-and-strains-stored-in-80c-5-sept-transformation-and-plating-attempt-2-samples-3414-working-to-obtain-more-strains-of-randomized-sample-3-6-sept-no-colonies-observed-8-sept-transformation-and-plating-attempt-4-samples-3414-9-sept-no-colonies-observed-11-sept-transformation-and-plating-attempt-5-samples-3414-slightly-reduced-antibiotic-concentration-12-sept-golden-gate-assembly-set-up-from-left-over-plasmid-for-3414-14-sept-transformation-and-plating-attempt-6-samples-3414-15-sept-1-colony-for-sample-3-none-for-4-and-14-17-sept-transformation-and-plating-attempt-7-samples-3414-18-sept-2-colonies-in-sample-3-none-for-4-and-14-21-sept-all-strains-13-variants-from-randomized-sample-3-11-model-generated-sequences-streaked-on-agar-plates-22-sept-colonies-picked-and-overnight-1ml-cultures-made-and-incubated-for-14-hrs-23-sept-fluorescence-assay-first-batch-of-results-obtained-25-sept-colonies-picked-from-streaked-plates-and-cultured-overnight-26-sept-plasmid-isolation-qiagen-kit-protocol-plasmid-concentration-obtained-were-very-poor-b3N-sub-5"
>
Colonies in all plates except samples 4 and 14. 10 colonies were observed in plate 3.
<br/>
Colonies were picked and cultured overnight. (2 cultures from each non-randomized sample, all 10 cultures of each colony from the randomized sample 3.
<br/>
</span>
<span
class=
"sept-transformation-plating-electroporation-was-done-for-all-the-samples-in-l-lactis-competent-cells-samples-were-incubated-for-1hr-after-which-they-were-spread-plated-and-incubated-for-24-hours-3-sept-colonies-in-all-plates-except-samples-4-and-14-10-colonies-were-observed-in-plate-3-colonies-were-picked-and-cultured-overnight-2-cultures-from-each-non-randomized-sample-all-10-cultures-of-each-colony-from-the-randomized-sample-3-4-sept-glycerol-added-and-strains-stored-in-80c-5-sept-transformation-and-plating-attempt-2-samples-3414-working-to-obtain-more-strains-of-randomized-sample-3-6-sept-no-colonies-observed-8-sept-transformation-and-plating-attempt-4-samples-3414-9-sept-no-colonies-observed-11-sept-transformation-and-plating-attempt-5-samples-3414-slightly-reduced-antibiotic-concentration-12-sept-golden-gate-assembly-set-up-from-left-over-plasmid-for-3414-14-sept-transformation-and-plating-attempt-6-samples-3414-15-sept-1-colony-for-sample-3-none-for-4-and-14-17-sept-transformation-and-plating-attempt-7-samples-3414-18-sept-2-colonies-in-sample-3-none-for-4-and-14-21-sept-all-strains-13-variants-from-randomized-sample-3-11-model-generated-sequences-streaked-on-agar-plates-22-sept-colonies-picked-and-overnight-1ml-cultures-made-and-incubated-for-14-hrs-23-sept-fluorescence-assay-first-batch-of-results-obtained-25-sept-colonies-picked-from-streaked-plates-and-cultured-overnight-26-sept-plasmid-isolation-qiagen-kit-protocol-plasmid-concentration-obtained-were-very-poor-b3N-sub-6"
>
<br/>
4 Sept
<br/>
</span>
<span
class=
"sept-transformation-plating-electroporation-was-done-for-all-the-samples-in-l-lactis-competent-cells-samples-were-incubated-for-1hr-after-which-they-were-spread-plated-and-incubated-for-24-hours-3-sept-colonies-in-all-plates-except-samples-4-and-14-10-colonies-were-observed-in-plate-3-colonies-were-picked-and-cultured-overnight-2-cultures-from-each-non-randomized-sample-all-10-cultures-of-each-colony-from-the-randomized-sample-3-4-sept-glycerol-added-and-strains-stored-in-80c-5-sept-transformation-and-plating-attempt-2-samples-3414-working-to-obtain-more-strains-of-randomized-sample-3-6-sept-no-colonies-observed-8-sept-transformation-and-plating-attempt-4-samples-3414-9-sept-no-colonies-observed-11-sept-transformation-and-plating-attempt-5-samples-3414-slightly-reduced-antibiotic-concentration-12-sept-golden-gate-assembly-set-up-from-left-over-plasmid-for-3414-14-sept-transformation-and-plating-attempt-6-samples-3414-15-sept-1-colony-for-sample-3-none-for-4-and-14-17-sept-transformation-and-plating-attempt-7-samples-3414-18-sept-2-colonies-in-sample-3-none-for-4-and-14-21-sept-all-strains-13-variants-from-randomized-sample-3-11-model-generated-sequences-streaked-on-agar-plates-22-sept-colonies-picked-and-overnight-1ml-cultures-made-and-incubated-for-14-hrs-23-sept-fluorescence-assay-first-batch-of-results-obtained-25-sept-colonies-picked-from-streaked-plates-and-cultured-overnight-26-sept-plasmid-isolation-qiagen-kit-protocol-plasmid-concentration-obtained-were-very-poor-b3N-sub-7"
>
Glycerol added and strains stored in -80C
<br/>
</span>
<span
class=
"sept-transformation-plating-electroporation-was-done-for-all-the-samples-in-l-lactis-competent-cells-samples-were-incubated-for-1hr-after-which-they-were-spread-plated-and-incubated-for-24-hours-3-sept-colonies-in-all-plates-except-samples-4-and-14-10-colonies-were-observed-in-plate-3-colonies-were-picked-and-cultured-overnight-2-cultures-from-each-non-randomized-sample-all-10-cultures-of-each-colony-from-the-randomized-sample-3-4-sept-glycerol-added-and-strains-stored-in-80c-5-sept-transformation-and-plating-attempt-2-samples-3414-working-to-obtain-more-strains-of-randomized-sample-3-6-sept-no-colonies-observed-8-sept-transformation-and-plating-attempt-4-samples-3414-9-sept-no-colonies-observed-11-sept-transformation-and-plating-attempt-5-samples-3414-slightly-reduced-antibiotic-concentration-12-sept-golden-gate-assembly-set-up-from-left-over-plasmid-for-3414-14-sept-transformation-and-plating-attempt-6-samples-3414-15-sept-1-colony-for-sample-3-none-for-4-and-14-17-sept-transformation-and-plating-attempt-7-samples-3414-18-sept-2-colonies-in-sample-3-none-for-4-and-14-21-sept-all-strains-13-variants-from-randomized-sample-3-11-model-generated-sequences-streaked-on-agar-plates-22-sept-colonies-picked-and-overnight-1ml-cultures-made-and-incubated-for-14-hrs-23-sept-fluorescence-assay-first-batch-of-results-obtained-25-sept-colonies-picked-from-streaked-plates-and-cultured-overnight-26-sept-plasmid-isolation-qiagen-kit-protocol-plasmid-concentration-obtained-were-very-poor-b3N-sub-8"
>
<br/>
5 Sept
<br/>
</span>
<span
class=
"sept-transformation-plating-electroporation-was-done-for-all-the-samples-in-l-lactis-competent-cells-samples-were-incubated-for-1hr-after-which-they-were-spread-plated-and-incubated-for-24-hours-3-sept-colonies-in-all-plates-except-samples-4-and-14-10-colonies-were-observed-in-plate-3-colonies-were-picked-and-cultured-overnight-2-cultures-from-each-non-randomized-sample-all-10-cultures-of-each-colony-from-the-randomized-sample-3-4-sept-glycerol-added-and-strains-stored-in-80c-5-sept-transformation-and-plating-attempt-2-samples-3414-working-to-obtain-more-strains-of-randomized-sample-3-6-sept-no-colonies-observed-8-sept-transformation-and-plating-attempt-4-samples-3414-9-sept-no-colonies-observed-11-sept-transformation-and-plating-attempt-5-samples-3414-slightly-reduced-antibiotic-concentration-12-sept-golden-gate-assembly-set-up-from-left-over-plasmid-for-3414-14-sept-transformation-and-plating-attempt-6-samples-3414-15-sept-1-colony-for-sample-3-none-for-4-and-14-17-sept-transformation-and-plating-attempt-7-samples-3414-18-sept-2-colonies-in-sample-3-none-for-4-and-14-21-sept-all-strains-13-variants-from-randomized-sample-3-11-model-generated-sequences-streaked-on-agar-plates-22-sept-colonies-picked-and-overnight-1ml-cultures-made-and-incubated-for-14-hrs-23-sept-fluorescence-assay-first-batch-of-results-obtained-25-sept-colonies-picked-from-streaked-plates-and-cultured-overnight-26-sept-plasmid-isolation-qiagen-kit-protocol-plasmid-concentration-obtained-were-very-poor-b3N-sub-9"
>
Transformation and plating (Attempt 2) (Samples 3,4,14) (Working to obtain more strains of randomized sample 3)
<br/>
<br/>
</span>
<span
class=
"sept-transformation-plating-electroporation-was-done-for-all-the-samples-in-l-lactis-competent-cells-samples-were-incubated-for-1hr-after-which-they-were-spread-plated-and-incubated-for-24-hours-3-sept-colonies-in-all-plates-except-samples-4-and-14-10-colonies-were-observed-in-plate-3-colonies-were-picked-and-cultured-overnight-2-cultures-from-each-non-randomized-sample-all-10-cultures-of-each-colony-from-the-randomized-sample-3-4-sept-glycerol-added-and-strains-stored-in-80c-5-sept-transformation-and-plating-attempt-2-samples-3414-working-to-obtain-more-strains-of-randomized-sample-3-6-sept-no-colonies-observed-8-sept-transformation-and-plating-attempt-4-samples-3414-9-sept-no-colonies-observed-11-sept-transformation-and-plating-attempt-5-samples-3414-slightly-reduced-antibiotic-concentration-12-sept-golden-gate-assembly-set-up-from-left-over-plasmid-for-3414-14-sept-transformation-and-plating-attempt-6-samples-3414-15-sept-1-colony-for-sample-3-none-for-4-and-14-17-sept-transformation-and-plating-attempt-7-samples-3414-18-sept-2-colonies-in-sample-3-none-for-4-and-14-21-sept-all-strains-13-variants-from-randomized-sample-3-11-model-generated-sequences-streaked-on-agar-plates-22-sept-colonies-picked-and-overnight-1ml-cultures-made-and-incubated-for-14-hrs-23-sept-fluorescence-assay-first-batch-of-results-obtained-25-sept-colonies-picked-from-streaked-plates-and-cultured-overnight-26-sept-plasmid-isolation-qiagen-kit-protocol-plasmid-concentration-obtained-were-very-poor-b3N-sub-10"
>
6 Sept
<br/>
</span>
<span
class=
"sept-transformation-plating-electroporation-was-done-for-all-the-samples-in-l-lactis-competent-cells-samples-were-incubated-for-1hr-after-which-they-were-spread-plated-and-incubated-for-24-hours-3-sept-colonies-in-all-plates-except-samples-4-and-14-10-colonies-were-observed-in-plate-3-colonies-were-picked-and-cultured-overnight-2-cultures-from-each-non-randomized-sample-all-10-cultures-of-each-colony-from-the-randomized-sample-3-4-sept-glycerol-added-and-strains-stored-in-80c-5-sept-transformation-and-plating-attempt-2-samples-3414-working-to-obtain-more-strains-of-randomized-sample-3-6-sept-no-colonies-observed-8-sept-transformation-and-plating-attempt-4-samples-3414-9-sept-no-colonies-observed-11-sept-transformation-and-plating-attempt-5-samples-3414-slightly-reduced-antibiotic-concentration-12-sept-golden-gate-assembly-set-up-from-left-over-plasmid-for-3414-14-sept-transformation-and-plating-attempt-6-samples-3414-15-sept-1-colony-for-sample-3-none-for-4-and-14-17-sept-transformation-and-plating-attempt-7-samples-3414-18-sept-2-colonies-in-sample-3-none-for-4-and-14-21-sept-all-strains-13-variants-from-randomized-sample-3-11-model-generated-sequences-streaked-on-agar-plates-22-sept-colonies-picked-and-overnight-1ml-cultures-made-and-incubated-for-14-hrs-23-sept-fluorescence-assay-first-batch-of-results-obtained-25-sept-colonies-picked-from-streaked-plates-and-cultured-overnight-26-sept-plasmid-isolation-qiagen-kit-protocol-plasmid-concentration-obtained-were-very-poor-b3N-sub-11"
>
No colonies observed
<br/>
<br/>
</span>
<span
class=
"sept-transformation-plating-electroporation-was-done-for-all-the-samples-in-l-lactis-competent-cells-samples-were-incubated-for-1hr-after-which-they-were-spread-plated-and-incubated-for-24-hours-3-sept-colonies-in-all-plates-except-samples-4-and-14-10-colonies-were-observed-in-plate-3-colonies-were-picked-and-cultured-overnight-2-cultures-from-each-non-randomized-sample-all-10-cultures-of-each-colony-from-the-randomized-sample-3-4-sept-glycerol-added-and-strains-stored-in-80c-5-sept-transformation-and-plating-attempt-2-samples-3414-working-to-obtain-more-strains-of-randomized-sample-3-6-sept-no-colonies-observed-8-sept-transformation-and-plating-attempt-4-samples-3414-9-sept-no-colonies-observed-11-sept-transformation-and-plating-attempt-5-samples-3414-slightly-reduced-antibiotic-concentration-12-sept-golden-gate-assembly-set-up-from-left-over-plasmid-for-3414-14-sept-transformation-and-plating-attempt-6-samples-3414-15-sept-1-colony-for-sample-3-none-for-4-and-14-17-sept-transformation-and-plating-attempt-7-samples-3414-18-sept-2-colonies-in-sample-3-none-for-4-and-14-21-sept-all-strains-13-variants-from-randomized-sample-3-11-model-generated-sequences-streaked-on-agar-plates-22-sept-colonies-picked-and-overnight-1ml-cultures-made-and-incubated-for-14-hrs-23-sept-fluorescence-assay-first-batch-of-results-obtained-25-sept-colonies-picked-from-streaked-plates-and-cultured-overnight-26-sept-plasmid-isolation-qiagen-kit-protocol-plasmid-concentration-obtained-were-very-poor-b3N-sub-12"
>
8 Sept
<br/>
</span>
<span
class=
"sept-transformation-plating-electroporation-was-done-for-all-the-samples-in-l-lactis-competent-cells-samples-were-incubated-for-1hr-after-which-they-were-spread-plated-and-incubated-for-24-hours-3-sept-colonies-in-all-plates-except-samples-4-and-14-10-colonies-were-observed-in-plate-3-colonies-were-picked-and-cultured-overnight-2-cultures-from-each-non-randomized-sample-all-10-cultures-of-each-colony-from-the-randomized-sample-3-4-sept-glycerol-added-and-strains-stored-in-80c-5-sept-transformation-and-plating-attempt-2-samples-3414-working-to-obtain-more-strains-of-randomized-sample-3-6-sept-no-colonies-observed-8-sept-transformation-and-plating-attempt-4-samples-3414-9-sept-no-colonies-observed-11-sept-transformation-and-plating-attempt-5-samples-3414-slightly-reduced-antibiotic-concentration-12-sept-golden-gate-assembly-set-up-from-left-over-plasmid-for-3414-14-sept-transformation-and-plating-attempt-6-samples-3414-15-sept-1-colony-for-sample-3-none-for-4-and-14-17-sept-transformation-and-plating-attempt-7-samples-3414-18-sept-2-colonies-in-sample-3-none-for-4-and-14-21-sept-all-strains-13-variants-from-randomized-sample-3-11-model-generated-sequences-streaked-on-agar-plates-22-sept-colonies-picked-and-overnight-1ml-cultures-made-and-incubated-for-14-hrs-23-sept-fluorescence-assay-first-batch-of-results-obtained-25-sept-colonies-picked-from-streaked-plates-and-cultured-overnight-26-sept-plasmid-isolation-qiagen-kit-protocol-plasmid-concentration-obtained-were-very-poor-b3N-sub-13"
>
Transformation and plating (Attempt 4) (Samples 3,4,14)
<br/>
<br/>
</span>
<span
class=
"sept-transformation-plating-electroporation-was-done-for-all-the-samples-in-l-lactis-competent-cells-samples-were-incubated-for-1hr-after-which-they-were-spread-plated-and-incubated-for-24-hours-3-sept-colonies-in-all-plates-except-samples-4-and-14-10-colonies-were-observed-in-plate-3-colonies-were-picked-and-cultured-overnight-2-cultures-from-each-non-randomized-sample-all-10-cultures-of-each-colony-from-the-randomized-sample-3-4-sept-glycerol-added-and-strains-stored-in-80c-5-sept-transformation-and-plating-attempt-2-samples-3414-working-to-obtain-more-strains-of-randomized-sample-3-6-sept-no-colonies-observed-8-sept-transformation-and-plating-attempt-4-samples-3414-9-sept-no-colonies-observed-11-sept-transformation-and-plating-attempt-5-samples-3414-slightly-reduced-antibiotic-concentration-12-sept-golden-gate-assembly-set-up-from-left-over-plasmid-for-3414-14-sept-transformation-and-plating-attempt-6-samples-3414-15-sept-1-colony-for-sample-3-none-for-4-and-14-17-sept-transformation-and-plating-attempt-7-samples-3414-18-sept-2-colonies-in-sample-3-none-for-4-and-14-21-sept-all-strains-13-variants-from-randomized-sample-3-11-model-generated-sequences-streaked-on-agar-plates-22-sept-colonies-picked-and-overnight-1ml-cultures-made-and-incubated-for-14-hrs-23-sept-fluorescence-assay-first-batch-of-results-obtained-25-sept-colonies-picked-from-streaked-plates-and-cultured-overnight-26-sept-plasmid-isolation-qiagen-kit-protocol-plasmid-concentration-obtained-were-very-poor-b3N-sub-14"
>
9 Sept
<br/>
</span>
<span
class=
"sept-transformation-plating-electroporation-was-done-for-all-the-samples-in-l-lactis-competent-cells-samples-were-incubated-for-1hr-after-which-they-were-spread-plated-and-incubated-for-24-hours-3-sept-colonies-in-all-plates-except-samples-4-and-14-10-colonies-were-observed-in-plate-3-colonies-were-picked-and-cultured-overnight-2-cultures-from-each-non-randomized-sample-all-10-cultures-of-each-colony-from-the-randomized-sample-3-4-sept-glycerol-added-and-strains-stored-in-80c-5-sept-transformation-and-plating-attempt-2-samples-3414-working-to-obtain-more-strains-of-randomized-sample-3-6-sept-no-colonies-observed-8-sept-transformation-and-plating-attempt-4-samples-3414-9-sept-no-colonies-observed-11-sept-transformation-and-plating-attempt-5-samples-3414-slightly-reduced-antibiotic-concentration-12-sept-golden-gate-assembly-set-up-from-left-over-plasmid-for-3414-14-sept-transformation-and-plating-attempt-6-samples-3414-15-sept-1-colony-for-sample-3-none-for-4-and-14-17-sept-transformation-and-plating-attempt-7-samples-3414-18-sept-2-colonies-in-sample-3-none-for-4-and-14-21-sept-all-strains-13-variants-from-randomized-sample-3-11-model-generated-sequences-streaked-on-agar-plates-22-sept-colonies-picked-and-overnight-1ml-cultures-made-and-incubated-for-14-hrs-23-sept-fluorescence-assay-first-batch-of-results-obtained-25-sept-colonies-picked-from-streaked-plates-and-cultured-overnight-26-sept-plasmid-isolation-qiagen-kit-protocol-plasmid-concentration-obtained-were-very-poor-b3N-sub-15"
>
No colonies observed
<br/>
<br/>
</span>
<span
class=
"sept-transformation-plating-electroporation-was-done-for-all-the-samples-in-l-lactis-competent-cells-samples-were-incubated-for-1hr-after-which-they-were-spread-plated-and-incubated-for-24-hours-3-sept-colonies-in-all-plates-except-samples-4-and-14-10-colonies-were-observed-in-plate-3-colonies-were-picked-and-cultured-overnight-2-cultures-from-each-non-randomized-sample-all-10-cultures-of-each-colony-from-the-randomized-sample-3-4-sept-glycerol-added-and-strains-stored-in-80c-5-sept-transformation-and-plating-attempt-2-samples-3414-working-to-obtain-more-strains-of-randomized-sample-3-6-sept-no-colonies-observed-8-sept-transformation-and-plating-attempt-4-samples-3414-9-sept-no-colonies-observed-11-sept-transformation-and-plating-attempt-5-samples-3414-slightly-reduced-antibiotic-concentration-12-sept-golden-gate-assembly-set-up-from-left-over-plasmid-for-3414-14-sept-transformation-and-plating-attempt-6-samples-3414-15-sept-1-colony-for-sample-3-none-for-4-and-14-17-sept-transformation-and-plating-attempt-7-samples-3414-18-sept-2-colonies-in-sample-3-none-for-4-and-14-21-sept-all-strains-13-variants-from-randomized-sample-3-11-model-generated-sequences-streaked-on-agar-plates-22-sept-colonies-picked-and-overnight-1ml-cultures-made-and-incubated-for-14-hrs-23-sept-fluorescence-assay-first-batch-of-results-obtained-25-sept-colonies-picked-from-streaked-plates-and-cultured-overnight-26-sept-plasmid-isolation-qiagen-kit-protocol-plasmid-concentration-obtained-were-very-poor-b3N-sub-16"
>
11 Sept
<br/>
</span>
<span
class=
"sept-transformation-plating-electroporation-was-done-for-all-the-samples-in-l-lactis-competent-cells-samples-were-incubated-for-1hr-after-which-they-were-spread-plated-and-incubated-for-24-hours-3-sept-colonies-in-all-plates-except-samples-4-and-14-10-colonies-were-observed-in-plate-3-colonies-were-picked-and-cultured-overnight-2-cultures-from-each-non-randomized-sample-all-10-cultures-of-each-colony-from-the-randomized-sample-3-4-sept-glycerol-added-and-strains-stored-in-80c-5-sept-transformation-and-plating-attempt-2-samples-3414-working-to-obtain-more-strains-of-randomized-sample-3-6-sept-no-colonies-observed-8-sept-transformation-and-plating-attempt-4-samples-3414-9-sept-no-colonies-observed-11-sept-transformation-and-plating-attempt-5-samples-3414-slightly-reduced-antibiotic-concentration-12-sept-golden-gate-assembly-set-up-from-left-over-plasmid-for-3414-14-sept-transformation-and-plating-attempt-6-samples-3414-15-sept-1-colony-for-sample-3-none-for-4-and-14-17-sept-transformation-and-plating-attempt-7-samples-3414-18-sept-2-colonies-in-sample-3-none-for-4-and-14-21-sept-all-strains-13-variants-from-randomized-sample-3-11-model-generated-sequences-streaked-on-agar-plates-22-sept-colonies-picked-and-overnight-1ml-cultures-made-and-incubated-for-14-hrs-23-sept-fluorescence-assay-first-batch-of-results-obtained-25-sept-colonies-picked-from-streaked-plates-and-cultured-overnight-26-sept-plasmid-isolation-qiagen-kit-protocol-plasmid-concentration-obtained-were-very-poor-b3N-sub-17"
>
Transformation and plating (Attempt 5) (Samples 3,4,14) (Slightly reduced antibiotic concentration)
<br/>
<br/>
</span>
<span
class=
"sept-transformation-plating-electroporation-was-done-for-all-the-samples-in-l-lactis-competent-cells-samples-were-incubated-for-1hr-after-which-they-were-spread-plated-and-incubated-for-24-hours-3-sept-colonies-in-all-plates-except-samples-4-and-14-10-colonies-were-observed-in-plate-3-colonies-were-picked-and-cultured-overnight-2-cultures-from-each-non-randomized-sample-all-10-cultures-of-each-colony-from-the-randomized-sample-3-4-sept-glycerol-added-and-strains-stored-in-80c-5-sept-transformation-and-plating-attempt-2-samples-3414-working-to-obtain-more-strains-of-randomized-sample-3-6-sept-no-colonies-observed-8-sept-transformation-and-plating-attempt-4-samples-3414-9-sept-no-colonies-observed-11-sept-transformation-and-plating-attempt-5-samples-3414-slightly-reduced-antibiotic-concentration-12-sept-golden-gate-assembly-set-up-from-left-over-plasmid-for-3414-14-sept-transformation-and-plating-attempt-6-samples-3414-15-sept-1-colony-for-sample-3-none-for-4-and-14-17-sept-transformation-and-plating-attempt-7-samples-3414-18-sept-2-colonies-in-sample-3-none-for-4-and-14-21-sept-all-strains-13-variants-from-randomized-sample-3-11-model-generated-sequences-streaked-on-agar-plates-22-sept-colonies-picked-and-overnight-1ml-cultures-made-and-incubated-for-14-hrs-23-sept-fluorescence-assay-first-batch-of-results-obtained-25-sept-colonies-picked-from-streaked-plates-and-cultured-overnight-26-sept-plasmid-isolation-qiagen-kit-protocol-plasmid-concentration-obtained-were-very-poor-b3N-sub-18"
>
12 Sept
<br/>
</span>
<span
class=
"sept-transformation-plating-electroporation-was-done-for-all-the-samples-in-l-lactis-competent-cells-samples-were-incubated-for-1hr-after-which-they-were-spread-plated-and-incubated-for-24-hours-3-sept-colonies-in-all-plates-except-samples-4-and-14-10-colonies-were-observed-in-plate-3-colonies-were-picked-and-cultured-overnight-2-cultures-from-each-non-randomized-sample-all-10-cultures-of-each-colony-from-the-randomized-sample-3-4-sept-glycerol-added-and-strains-stored-in-80c-5-sept-transformation-and-plating-attempt-2-samples-3414-working-to-obtain-more-strains-of-randomized-sample-3-6-sept-no-colonies-observed-8-sept-transformation-and-plating-attempt-4-samples-3414-9-sept-no-colonies-observed-11-sept-transformation-and-plating-attempt-5-samples-3414-slightly-reduced-antibiotic-concentration-12-sept-golden-gate-assembly-set-up-from-left-over-plasmid-for-3414-14-sept-transformation-and-plating-attempt-6-samples-3414-15-sept-1-colony-for-sample-3-none-for-4-and-14-17-sept-transformation-and-plating-attempt-7-samples-3414-18-sept-2-colonies-in-sample-3-none-for-4-and-14-21-sept-all-strains-13-variants-from-randomized-sample-3-11-model-generated-sequences-streaked-on-agar-plates-22-sept-colonies-picked-and-overnight-1ml-cultures-made-and-incubated-for-14-hrs-23-sept-fluorescence-assay-first-batch-of-results-obtained-25-sept-colonies-picked-from-streaked-plates-and-cultured-overnight-26-sept-plasmid-isolation-qiagen-kit-protocol-plasmid-concentration-obtained-were-very-poor-b3N-sub-19"
>
Golden Gate Assembly set up from left over plasmid for 3,4,14
<br/>
<br/>
</span>
<span
class=
"sept-transformation-plating-electroporation-was-done-for-all-the-samples-in-l-lactis-competent-cells-samples-were-incubated-for-1hr-after-which-they-were-spread-plated-and-incubated-for-24-hours-3-sept-colonies-in-all-plates-except-samples-4-and-14-10-colonies-were-observed-in-plate-3-colonies-were-picked-and-cultured-overnight-2-cultures-from-each-non-randomized-sample-all-10-cultures-of-each-colony-from-the-randomized-sample-3-4-sept-glycerol-added-and-strains-stored-in-80c-5-sept-transformation-and-plating-attempt-2-samples-3414-working-to-obtain-more-strains-of-randomized-sample-3-6-sept-no-colonies-observed-8-sept-transformation-and-plating-attempt-4-samples-3414-9-sept-no-colonies-observed-11-sept-transformation-and-plating-attempt-5-samples-3414-slightly-reduced-antibiotic-concentration-12-sept-golden-gate-assembly-set-up-from-left-over-plasmid-for-3414-14-sept-transformation-and-plating-attempt-6-samples-3414-15-sept-1-colony-for-sample-3-none-for-4-and-14-17-sept-transformation-and-plating-attempt-7-samples-3414-18-sept-2-colonies-in-sample-3-none-for-4-and-14-21-sept-all-strains-13-variants-from-randomized-sample-3-11-model-generated-sequences-streaked-on-agar-plates-22-sept-colonies-picked-and-overnight-1ml-cultures-made-and-incubated-for-14-hrs-23-sept-fluorescence-assay-first-batch-of-results-obtained-25-sept-colonies-picked-from-streaked-plates-and-cultured-overnight-26-sept-plasmid-isolation-qiagen-kit-protocol-plasmid-concentration-obtained-were-very-poor-b3N-sub-20"
>
14 Sept
<br/>
</span>
<span
class=
"sept-transformation-plating-electroporation-was-done-for-all-the-samples-in-l-lactis-competent-cells-samples-were-incubated-for-1hr-after-which-they-were-spread-plated-and-incubated-for-24-hours-3-sept-colonies-in-all-plates-except-samples-4-and-14-10-colonies-were-observed-in-plate-3-colonies-were-picked-and-cultured-overnight-2-cultures-from-each-non-randomized-sample-all-10-cultures-of-each-colony-from-the-randomized-sample-3-4-sept-glycerol-added-and-strains-stored-in-80c-5-sept-transformation-and-plating-attempt-2-samples-3414-working-to-obtain-more-strains-of-randomized-sample-3-6-sept-no-colonies-observed-8-sept-transformation-and-plating-attempt-4-samples-3414-9-sept-no-colonies-observed-11-sept-transformation-and-plating-attempt-5-samples-3414-slightly-reduced-antibiotic-concentration-12-sept-golden-gate-assembly-set-up-from-left-over-plasmid-for-3414-14-sept-transformation-and-plating-attempt-6-samples-3414-15-sept-1-colony-for-sample-3-none-for-4-and-14-17-sept-transformation-and-plating-attempt-7-samples-3414-18-sept-2-colonies-in-sample-3-none-for-4-and-14-21-sept-all-strains-13-variants-from-randomized-sample-3-11-model-generated-sequences-streaked-on-agar-plates-22-sept-colonies-picked-and-overnight-1ml-cultures-made-and-incubated-for-14-hrs-23-sept-fluorescence-assay-first-batch-of-results-obtained-25-sept-colonies-picked-from-streaked-plates-and-cultured-overnight-26-sept-plasmid-isolation-qiagen-kit-protocol-plasmid-concentration-obtained-were-very-poor-b3N-sub-21"
>
Transformation and plating (Attempt 6) (Samples 3,4,14)
<br/>
<br/>
</span>
<span
class=
"sept-transformation-plating-electroporation-was-done-for-all-the-samples-in-l-lactis-competent-cells-samples-were-incubated-for-1hr-after-which-they-were-spread-plated-and-incubated-for-24-hours-3-sept-colonies-in-all-plates-except-samples-4-and-14-10-colonies-were-observed-in-plate-3-colonies-were-picked-and-cultured-overnight-2-cultures-from-each-non-randomized-sample-all-10-cultures-of-each-colony-from-the-randomized-sample-3-4-sept-glycerol-added-and-strains-stored-in-80c-5-sept-transformation-and-plating-attempt-2-samples-3414-working-to-obtain-more-strains-of-randomized-sample-3-6-sept-no-colonies-observed-8-sept-transformation-and-plating-attempt-4-samples-3414-9-sept-no-colonies-observed-11-sept-transformation-and-plating-attempt-5-samples-3414-slightly-reduced-antibiotic-concentration-12-sept-golden-gate-assembly-set-up-from-left-over-plasmid-for-3414-14-sept-transformation-and-plating-attempt-6-samples-3414-15-sept-1-colony-for-sample-3-none-for-4-and-14-17-sept-transformation-and-plating-attempt-7-samples-3414-18-sept-2-colonies-in-sample-3-none-for-4-and-14-21-sept-all-strains-13-variants-from-randomized-sample-3-11-model-generated-sequences-streaked-on-agar-plates-22-sept-colonies-picked-and-overnight-1ml-cultures-made-and-incubated-for-14-hrs-23-sept-fluorescence-assay-first-batch-of-results-obtained-25-sept-colonies-picked-from-streaked-plates-and-cultured-overnight-26-sept-plasmid-isolation-qiagen-kit-protocol-plasmid-concentration-obtained-were-very-poor-b3N-sub-22"
>
15 Sept
<br/>
</span>
<span
class=
"sept-transformation-plating-electroporation-was-done-for-all-the-samples-in-l-lactis-competent-cells-samples-were-incubated-for-1hr-after-which-they-were-spread-plated-and-incubated-for-24-hours-3-sept-colonies-in-all-plates-except-samples-4-and-14-10-colonies-were-observed-in-plate-3-colonies-were-picked-and-cultured-overnight-2-cultures-from-each-non-randomized-sample-all-10-cultures-of-each-colony-from-the-randomized-sample-3-4-sept-glycerol-added-and-strains-stored-in-80c-5-sept-transformation-and-plating-attempt-2-samples-3414-working-to-obtain-more-strains-of-randomized-sample-3-6-sept-no-colonies-observed-8-sept-transformation-and-plating-attempt-4-samples-3414-9-sept-no-colonies-observed-11-sept-transformation-and-plating-attempt-5-samples-3414-slightly-reduced-antibiotic-concentration-12-sept-golden-gate-assembly-set-up-from-left-over-plasmid-for-3414-14-sept-transformation-and-plating-attempt-6-samples-3414-15-sept-1-colony-for-sample-3-none-for-4-and-14-17-sept-transformation-and-plating-attempt-7-samples-3414-18-sept-2-colonies-in-sample-3-none-for-4-and-14-21-sept-all-strains-13-variants-from-randomized-sample-3-11-model-generated-sequences-streaked-on-agar-plates-22-sept-colonies-picked-and-overnight-1ml-cultures-made-and-incubated-for-14-hrs-23-sept-fluorescence-assay-first-batch-of-results-obtained-25-sept-colonies-picked-from-streaked-plates-and-cultured-overnight-26-sept-plasmid-isolation-qiagen-kit-protocol-plasmid-concentration-obtained-were-very-poor-b3N-sub-23"
>
1 colony for sample 3. None for 4 and 14.
<br/>
<br/>
</span>
<span
class=
"sept-transformation-plating-electroporation-was-done-for-all-the-samples-in-l-lactis-competent-cells-samples-were-incubated-for-1hr-after-which-they-were-spread-plated-and-incubated-for-24-hours-3-sept-colonies-in-all-plates-except-samples-4-and-14-10-colonies-were-observed-in-plate-3-colonies-were-picked-and-cultured-overnight-2-cultures-from-each-non-randomized-sample-all-10-cultures-of-each-colony-from-the-randomized-sample-3-4-sept-glycerol-added-and-strains-stored-in-80c-5-sept-transformation-and-plating-attempt-2-samples-3414-working-to-obtain-more-strains-of-randomized-sample-3-6-sept-no-colonies-observed-8-sept-transformation-and-plating-attempt-4-samples-3414-9-sept-no-colonies-observed-11-sept-transformation-and-plating-attempt-5-samples-3414-slightly-reduced-antibiotic-concentration-12-sept-golden-gate-assembly-set-up-from-left-over-plasmid-for-3414-14-sept-transformation-and-plating-attempt-6-samples-3414-15-sept-1-colony-for-sample-3-none-for-4-and-14-17-sept-transformation-and-plating-attempt-7-samples-3414-18-sept-2-colonies-in-sample-3-none-for-4-and-14-21-sept-all-strains-13-variants-from-randomized-sample-3-11-model-generated-sequences-streaked-on-agar-plates-22-sept-colonies-picked-and-overnight-1ml-cultures-made-and-incubated-for-14-hrs-23-sept-fluorescence-assay-first-batch-of-results-obtained-25-sept-colonies-picked-from-streaked-plates-and-cultured-overnight-26-sept-plasmid-isolation-qiagen-kit-protocol-plasmid-concentration-obtained-were-very-poor-b3N-sub-24"
>
17 Sept
<br/>
</span>
<span
class=
"sept-transformation-plating-electroporation-was-done-for-all-the-samples-in-l-lactis-competent-cells-samples-were-incubated-for-1hr-after-which-they-were-spread-plated-and-incubated-for-24-hours-3-sept-colonies-in-all-plates-except-samples-4-and-14-10-colonies-were-observed-in-plate-3-colonies-were-picked-and-cultured-overnight-2-cultures-from-each-non-randomized-sample-all-10-cultures-of-each-colony-from-the-randomized-sample-3-4-sept-glycerol-added-and-strains-stored-in-80c-5-sept-transformation-and-plating-attempt-2-samples-3414-working-to-obtain-more-strains-of-randomized-sample-3-6-sept-no-colonies-observed-8-sept-transformation-and-plating-attempt-4-samples-3414-9-sept-no-colonies-observed-11-sept-transformation-and-plating-attempt-5-samples-3414-slightly-reduced-antibiotic-concentration-12-sept-golden-gate-assembly-set-up-from-left-over-plasmid-for-3414-14-sept-transformation-and-plating-attempt-6-samples-3414-15-sept-1-colony-for-sample-3-none-for-4-and-14-17-sept-transformation-and-plating-attempt-7-samples-3414-18-sept-2-colonies-in-sample-3-none-for-4-and-14-21-sept-all-strains-13-variants-from-randomized-sample-3-11-model-generated-sequences-streaked-on-agar-plates-22-sept-colonies-picked-and-overnight-1ml-cultures-made-and-incubated-for-14-hrs-23-sept-fluorescence-assay-first-batch-of-results-obtained-25-sept-colonies-picked-from-streaked-plates-and-cultured-overnight-26-sept-plasmid-isolation-qiagen-kit-protocol-plasmid-concentration-obtained-were-very-poor-b3N-sub-25"
>
Transformation and plating (Attempt 7) (Samples 3,4,14)
<br/>
<br/>
</span>
<span
class=
"sept-transformation-plating-electroporation-was-done-for-all-the-samples-in-l-lactis-competent-cells-samples-were-incubated-for-1hr-after-which-they-were-spread-plated-and-incubated-for-24-hours-3-sept-colonies-in-all-plates-except-samples-4-and-14-10-colonies-were-observed-in-plate-3-colonies-were-picked-and-cultured-overnight-2-cultures-from-each-non-randomized-sample-all-10-cultures-of-each-colony-from-the-randomized-sample-3-4-sept-glycerol-added-and-strains-stored-in-80c-5-sept-transformation-and-plating-attempt-2-samples-3414-working-to-obtain-more-strains-of-randomized-sample-3-6-sept-no-colonies-observed-8-sept-transformation-and-plating-attempt-4-samples-3414-9-sept-no-colonies-observed-11-sept-transformation-and-plating-attempt-5-samples-3414-slightly-reduced-antibiotic-concentration-12-sept-golden-gate-assembly-set-up-from-left-over-plasmid-for-3414-14-sept-transformation-and-plating-attempt-6-samples-3414-15-sept-1-colony-for-sample-3-none-for-4-and-14-17-sept-transformation-and-plating-attempt-7-samples-3414-18-sept-2-colonies-in-sample-3-none-for-4-and-14-21-sept-all-strains-13-variants-from-randomized-sample-3-11-model-generated-sequences-streaked-on-agar-plates-22-sept-colonies-picked-and-overnight-1ml-cultures-made-and-incubated-for-14-hrs-23-sept-fluorescence-assay-first-batch-of-results-obtained-25-sept-colonies-picked-from-streaked-plates-and-cultured-overnight-26-sept-plasmid-isolation-qiagen-kit-protocol-plasmid-concentration-obtained-were-very-poor-b3N-sub-26"
>
18 Sept
<br/>
</span>
<span
class=
"sept-transformation-plating-electroporation-was-done-for-all-the-samples-in-l-lactis-competent-cells-samples-were-incubated-for-1hr-after-which-they-were-spread-plated-and-incubated-for-24-hours-3-sept-colonies-in-all-plates-except-samples-4-and-14-10-colonies-were-observed-in-plate-3-colonies-were-picked-and-cultured-overnight-2-cultures-from-each-non-randomized-sample-all-10-cultures-of-each-colony-from-the-randomized-sample-3-4-sept-glycerol-added-and-strains-stored-in-80c-5-sept-transformation-and-plating-attempt-2-samples-3414-working-to-obtain-more-strains-of-randomized-sample-3-6-sept-no-colonies-observed-8-sept-transformation-and-plating-attempt-4-samples-3414-9-sept-no-colonies-observed-11-sept-transformation-and-plating-attempt-5-samples-3414-slightly-reduced-antibiotic-concentration-12-sept-golden-gate-assembly-set-up-from-left-over-plasmid-for-3414-14-sept-transformation-and-plating-attempt-6-samples-3414-15-sept-1-colony-for-sample-3-none-for-4-and-14-17-sept-transformation-and-plating-attempt-7-samples-3414-18-sept-2-colonies-in-sample-3-none-for-4-and-14-21-sept-all-strains-13-variants-from-randomized-sample-3-11-model-generated-sequences-streaked-on-agar-plates-22-sept-colonies-picked-and-overnight-1ml-cultures-made-and-incubated-for-14-hrs-23-sept-fluorescence-assay-first-batch-of-results-obtained-25-sept-colonies-picked-from-streaked-plates-and-cultured-overnight-26-sept-plasmid-isolation-qiagen-kit-protocol-plasmid-concentration-obtained-were-very-poor-b3N-sub-27"
>
2 colonies in sample 3. None for 4 and 14.
<br/>
<br/>
</span>
<span
class=
"sept-transformation-plating-electroporation-was-done-for-all-the-samples-in-l-lactis-competent-cells-samples-were-incubated-for-1hr-after-which-they-were-spread-plated-and-incubated-for-24-hours-3-sept-colonies-in-all-plates-except-samples-4-and-14-10-colonies-were-observed-in-plate-3-colonies-were-picked-and-cultured-overnight-2-cultures-from-each-non-randomized-sample-all-10-cultures-of-each-colony-from-the-randomized-sample-3-4-sept-glycerol-added-and-strains-stored-in-80c-5-sept-transformation-and-plating-attempt-2-samples-3414-working-to-obtain-more-strains-of-randomized-sample-3-6-sept-no-colonies-observed-8-sept-transformation-and-plating-attempt-4-samples-3414-9-sept-no-colonies-observed-11-sept-transformation-and-plating-attempt-5-samples-3414-slightly-reduced-antibiotic-concentration-12-sept-golden-gate-assembly-set-up-from-left-over-plasmid-for-3414-14-sept-transformation-and-plating-attempt-6-samples-3414-15-sept-1-colony-for-sample-3-none-for-4-and-14-17-sept-transformation-and-plating-attempt-7-samples-3414-18-sept-2-colonies-in-sample-3-none-for-4-and-14-21-sept-all-strains-13-variants-from-randomized-sample-3-11-model-generated-sequences-streaked-on-agar-plates-22-sept-colonies-picked-and-overnight-1ml-cultures-made-and-incubated-for-14-hrs-23-sept-fluorescence-assay-first-batch-of-results-obtained-25-sept-colonies-picked-from-streaked-plates-and-cultured-overnight-26-sept-plasmid-isolation-qiagen-kit-protocol-plasmid-concentration-obtained-were-very-poor-b3N-sub-28"
>
21 Sept
<br/>
</span>
<span
class=
"sept-transformation-plating-electroporation-was-done-for-all-the-samples-in-l-lactis-competent-cells-samples-were-incubated-for-1hr-after-which-they-were-spread-plated-and-incubated-for-24-hours-3-sept-colonies-in-all-plates-except-samples-4-and-14-10-colonies-were-observed-in-plate-3-colonies-were-picked-and-cultured-overnight-2-cultures-from-each-non-randomized-sample-all-10-cultures-of-each-colony-from-the-randomized-sample-3-4-sept-glycerol-added-and-strains-stored-in-80c-5-sept-transformation-and-plating-attempt-2-samples-3414-working-to-obtain-more-strains-of-randomized-sample-3-6-sept-no-colonies-observed-8-sept-transformation-and-plating-attempt-4-samples-3414-9-sept-no-colonies-observed-11-sept-transformation-and-plating-attempt-5-samples-3414-slightly-reduced-antibiotic-concentration-12-sept-golden-gate-assembly-set-up-from-left-over-plasmid-for-3414-14-sept-transformation-and-plating-attempt-6-samples-3414-15-sept-1-colony-for-sample-3-none-for-4-and-14-17-sept-transformation-and-plating-attempt-7-samples-3414-18-sept-2-colonies-in-sample-3-none-for-4-and-14-21-sept-all-strains-13-variants-from-randomized-sample-3-11-model-generated-sequences-streaked-on-agar-plates-22-sept-colonies-picked-and-overnight-1ml-cultures-made-and-incubated-for-14-hrs-23-sept-fluorescence-assay-first-batch-of-results-obtained-25-sept-colonies-picked-from-streaked-plates-and-cultured-overnight-26-sept-plasmid-isolation-qiagen-kit-protocol-plasmid-concentration-obtained-were-very-poor-b3N-sub-29"
>
All strains (13 variants from randomized sample 3, 11 model generated sequences) streaked on agar plates
<br/>
<br/>
</span>
<span
class=
"sept-transformation-plating-electroporation-was-done-for-all-the-samples-in-l-lactis-competent-cells-samples-were-incubated-for-1hr-after-which-they-were-spread-plated-and-incubated-for-24-hours-3-sept-colonies-in-all-plates-except-samples-4-and-14-10-colonies-were-observed-in-plate-3-colonies-were-picked-and-cultured-overnight-2-cultures-from-each-non-randomized-sample-all-10-cultures-of-each-colony-from-the-randomized-sample-3-4-sept-glycerol-added-and-strains-stored-in-80c-5-sept-transformation-and-plating-attempt-2-samples-3414-working-to-obtain-more-strains-of-randomized-sample-3-6-sept-no-colonies-observed-8-sept-transformation-and-plating-attempt-4-samples-3414-9-sept-no-colonies-observed-11-sept-transformation-and-plating-attempt-5-samples-3414-slightly-reduced-antibiotic-concentration-12-sept-golden-gate-assembly-set-up-from-left-over-plasmid-for-3414-14-sept-transformation-and-plating-attempt-6-samples-3414-15-sept-1-colony-for-sample-3-none-for-4-and-14-17-sept-transformation-and-plating-attempt-7-samples-3414-18-sept-2-colonies-in-sample-3-none-for-4-and-14-21-sept-all-strains-13-variants-from-randomized-sample-3-11-model-generated-sequences-streaked-on-agar-plates-22-sept-colonies-picked-and-overnight-1ml-cultures-made-and-incubated-for-14-hrs-23-sept-fluorescence-assay-first-batch-of-results-obtained-25-sept-colonies-picked-from-streaked-plates-and-cultured-overnight-26-sept-plasmid-isolation-qiagen-kit-protocol-plasmid-concentration-obtained-were-very-poor-b3N-sub-30"
>
22 Sept
<br/>
</span>
<span
class=
"sept-transformation-plating-electroporation-was-done-for-all-the-samples-in-l-lactis-competent-cells-samples-were-incubated-for-1hr-after-which-they-were-spread-plated-and-incubated-for-24-hours-3-sept-colonies-in-all-plates-except-samples-4-and-14-10-colonies-were-observed-in-plate-3-colonies-were-picked-and-cultured-overnight-2-cultures-from-each-non-randomized-sample-all-10-cultures-of-each-colony-from-the-randomized-sample-3-4-sept-glycerol-added-and-strains-stored-in-80c-5-sept-transformation-and-plating-attempt-2-samples-3414-working-to-obtain-more-strains-of-randomized-sample-3-6-sept-no-colonies-observed-8-sept-transformation-and-plating-attempt-4-samples-3414-9-sept-no-colonies-observed-11-sept-transformation-and-plating-attempt-5-samples-3414-slightly-reduced-antibiotic-concentration-12-sept-golden-gate-assembly-set-up-from-left-over-plasmid-for-3414-14-sept-transformation-and-plating-attempt-6-samples-3414-15-sept-1-colony-for-sample-3-none-for-4-and-14-17-sept-transformation-and-plating-attempt-7-samples-3414-18-sept-2-colonies-in-sample-3-none-for-4-and-14-21-sept-all-strains-13-variants-from-randomized-sample-3-11-model-generated-sequences-streaked-on-agar-plates-22-sept-colonies-picked-and-overnight-1ml-cultures-made-and-incubated-for-14-hrs-23-sept-fluorescence-assay-first-batch-of-results-obtained-25-sept-colonies-picked-from-streaked-plates-and-cultured-overnight-26-sept-plasmid-isolation-qiagen-kit-protocol-plasmid-concentration-obtained-were-very-poor-b3N-sub-31"
>
Colonies picked and Overnight 1ml cultures made and incubated for 14 hrs
<br/>
<br/>
</span>
<span
class=
"sept-transformation-plating-electroporation-was-done-for-all-the-samples-in-l-lactis-competent-cells-samples-were-incubated-for-1hr-after-which-they-were-spread-plated-and-incubated-for-24-hours-3-sept-colonies-in-all-plates-except-samples-4-and-14-10-colonies-were-observed-in-plate-3-colonies-were-picked-and-cultured-overnight-2-cultures-from-each-non-randomized-sample-all-10-cultures-of-each-colony-from-the-randomized-sample-3-4-sept-glycerol-added-and-strains-stored-in-80c-5-sept-transformation-and-plating-attempt-2-samples-3414-working-to-obtain-more-strains-of-randomized-sample-3-6-sept-no-colonies-observed-8-sept-transformation-and-plating-attempt-4-samples-3414-9-sept-no-colonies-observed-11-sept-transformation-and-plating-attempt-5-samples-3414-slightly-reduced-antibiotic-concentration-12-sept-golden-gate-assembly-set-up-from-left-over-plasmid-for-3414-14-sept-transformation-and-plating-attempt-6-samples-3414-15-sept-1-colony-for-sample-3-none-for-4-and-14-17-sept-transformation-and-plating-attempt-7-samples-3414-18-sept-2-colonies-in-sample-3-none-for-4-and-14-21-sept-all-strains-13-variants-from-randomized-sample-3-11-model-generated-sequences-streaked-on-agar-plates-22-sept-colonies-picked-and-overnight-1ml-cultures-made-and-incubated-for-14-hrs-23-sept-fluorescence-assay-first-batch-of-results-obtained-25-sept-colonies-picked-from-streaked-plates-and-cultured-overnight-26-sept-plasmid-isolation-qiagen-kit-protocol-plasmid-concentration-obtained-were-very-poor-b3N-sub-32"
>
23 Sept
<br/>
</span>
<span
class=
"sept-transformation-plating-electroporation-was-done-for-all-the-samples-in-l-lactis-competent-cells-samples-were-incubated-for-1hr-after-which-they-were-spread-plated-and-incubated-for-24-hours-3-sept-colonies-in-all-plates-except-samples-4-and-14-10-colonies-were-observed-in-plate-3-colonies-were-picked-and-cultured-overnight-2-cultures-from-each-non-randomized-sample-all-10-cultures-of-each-colony-from-the-randomized-sample-3-4-sept-glycerol-added-and-strains-stored-in-80c-5-sept-transformation-and-plating-attempt-2-samples-3414-working-to-obtain-more-strains-of-randomized-sample-3-6-sept-no-colonies-observed-8-sept-transformation-and-plating-attempt-4-samples-3414-9-sept-no-colonies-observed-11-sept-transformation-and-plating-attempt-5-samples-3414-slightly-reduced-antibiotic-concentration-12-sept-golden-gate-assembly-set-up-from-left-over-plasmid-for-3414-14-sept-transformation-and-plating-attempt-6-samples-3414-15-sept-1-colony-for-sample-3-none-for-4-and-14-17-sept-transformation-and-plating-attempt-7-samples-3414-18-sept-2-colonies-in-sample-3-none-for-4-and-14-21-sept-all-strains-13-variants-from-randomized-sample-3-11-model-generated-sequences-streaked-on-agar-plates-22-sept-colonies-picked-and-overnight-1ml-cultures-made-and-incubated-for-14-hrs-23-sept-fluorescence-assay-first-batch-of-results-obtained-25-sept-colonies-picked-from-streaked-plates-and-cultured-overnight-26-sept-plasmid-isolation-qiagen-kit-protocol-plasmid-concentration-obtained-were-very-poor-b3N-sub-33"
>
Fluorescence Assay
<br/>
</span>
<span
class=
"sept-transformation-plating-electroporation-was-done-for-all-the-samples-in-l-lactis-competent-cells-samples-were-incubated-for-1hr-after-which-they-were-spread-plated-and-incubated-for-24-hours-3-sept-colonies-in-all-plates-except-samples-4-and-14-10-colonies-were-observed-in-plate-3-colonies-were-picked-and-cultured-overnight-2-cultures-from-each-non-randomized-sample-all-10-cultures-of-each-colony-from-the-randomized-sample-3-4-sept-glycerol-added-and-strains-stored-in-80c-5-sept-transformation-and-plating-attempt-2-samples-3414-working-to-obtain-more-strains-of-randomized-sample-3-6-sept-no-colonies-observed-8-sept-transformation-and-plating-attempt-4-samples-3414-9-sept-no-colonies-observed-11-sept-transformation-and-plating-attempt-5-samples-3414-slightly-reduced-antibiotic-concentration-12-sept-golden-gate-assembly-set-up-from-left-over-plasmid-for-3414-14-sept-transformation-and-plating-attempt-6-samples-3414-15-sept-1-colony-for-sample-3-none-for-4-and-14-17-sept-transformation-and-plating-attempt-7-samples-3414-18-sept-2-colonies-in-sample-3-none-for-4-and-14-21-sept-all-strains-13-variants-from-randomized-sample-3-11-model-generated-sequences-streaked-on-agar-plates-22-sept-colonies-picked-and-overnight-1ml-cultures-made-and-incubated-for-14-hrs-23-sept-fluorescence-assay-first-batch-of-results-obtained-25-sept-colonies-picked-from-streaked-plates-and-cultured-overnight-26-sept-plasmid-isolation-qiagen-kit-protocol-plasmid-concentration-obtained-were-very-poor-b3N-sub-34"
>
First batch of results obtained.
<br/>
</span>
<span
class=
"sept-transformation-plating-electroporation-was-done-for-all-the-samples-in-l-lactis-competent-cells-samples-were-incubated-for-1hr-after-which-they-were-spread-plated-and-incubated-for-24-hours-3-sept-colonies-in-all-plates-except-samples-4-and-14-10-colonies-were-observed-in-plate-3-colonies-were-picked-and-cultured-overnight-2-cultures-from-each-non-randomized-sample-all-10-cultures-of-each-colony-from-the-randomized-sample-3-4-sept-glycerol-added-and-strains-stored-in-80c-5-sept-transformation-and-plating-attempt-2-samples-3414-working-to-obtain-more-strains-of-randomized-sample-3-6-sept-no-colonies-observed-8-sept-transformation-and-plating-attempt-4-samples-3414-9-sept-no-colonies-observed-11-sept-transformation-and-plating-attempt-5-samples-3414-slightly-reduced-antibiotic-concentration-12-sept-golden-gate-assembly-set-up-from-left-over-plasmid-for-3414-14-sept-transformation-and-plating-attempt-6-samples-3414-15-sept-1-colony-for-sample-3-none-for-4-and-14-17-sept-transformation-and-plating-attempt-7-samples-3414-18-sept-2-colonies-in-sample-3-none-for-4-and-14-21-sept-all-strains-13-variants-from-randomized-sample-3-11-model-generated-sequences-streaked-on-agar-plates-22-sept-colonies-picked-and-overnight-1ml-cultures-made-and-incubated-for-14-hrs-23-sept-fluorescence-assay-first-batch-of-results-obtained-25-sept-colonies-picked-from-streaked-plates-and-cultured-overnight-26-sept-plasmid-isolation-qiagen-kit-protocol-plasmid-concentration-obtained-were-very-poor-b3N-sub-35"
>
<br/>
</span>
<span
class=
"sept-transformation-plating-electroporation-was-done-for-all-the-samples-in-l-lactis-competent-cells-samples-were-incubated-for-1hr-after-which-they-were-spread-plated-and-incubated-for-24-hours-3-sept-colonies-in-all-plates-except-samples-4-and-14-10-colonies-were-observed-in-plate-3-colonies-were-picked-and-cultured-overnight-2-cultures-from-each-non-randomized-sample-all-10-cultures-of-each-colony-from-the-randomized-sample-3-4-sept-glycerol-added-and-strains-stored-in-80c-5-sept-transformation-and-plating-attempt-2-samples-3414-working-to-obtain-more-strains-of-randomized-sample-3-6-sept-no-colonies-observed-8-sept-transformation-and-plating-attempt-4-samples-3414-9-sept-no-colonies-observed-11-sept-transformation-and-plating-attempt-5-samples-3414-slightly-reduced-antibiotic-concentration-12-sept-golden-gate-assembly-set-up-from-left-over-plasmid-for-3414-14-sept-transformation-and-plating-attempt-6-samples-3414-15-sept-1-colony-for-sample-3-none-for-4-and-14-17-sept-transformation-and-plating-attempt-7-samples-3414-18-sept-2-colonies-in-sample-3-none-for-4-and-14-21-sept-all-strains-13-variants-from-randomized-sample-3-11-model-generated-sequences-streaked-on-agar-plates-22-sept-colonies-picked-and-overnight-1ml-cultures-made-and-incubated-for-14-hrs-23-sept-fluorescence-assay-first-batch-of-results-obtained-25-sept-colonies-picked-from-streaked-plates-and-cultured-overnight-26-sept-plasmid-isolation-qiagen-kit-protocol-plasmid-concentration-obtained-were-very-poor-b3N-sub-36"
>
25 Sept
<br/>
</span>
<span
class=
"sept-transformation-plating-electroporation-was-done-for-all-the-samples-in-l-lactis-competent-cells-samples-were-incubated-for-1hr-after-which-they-were-spread-plated-and-incubated-for-24-hours-3-sept-colonies-in-all-plates-except-samples-4-and-14-10-colonies-were-observed-in-plate-3-colonies-were-picked-and-cultured-overnight-2-cultures-from-each-non-randomized-sample-all-10-cultures-of-each-colony-from-the-randomized-sample-3-4-sept-glycerol-added-and-strains-stored-in-80c-5-sept-transformation-and-plating-attempt-2-samples-3414-working-to-obtain-more-strains-of-randomized-sample-3-6-sept-no-colonies-observed-8-sept-transformation-and-plating-attempt-4-samples-3414-9-sept-no-colonies-observed-11-sept-transformation-and-plating-attempt-5-samples-3414-slightly-reduced-antibiotic-concentration-12-sept-golden-gate-assembly-set-up-from-left-over-plasmid-for-3414-14-sept-transformation-and-plating-attempt-6-samples-3414-15-sept-1-colony-for-sample-3-none-for-4-and-14-17-sept-transformation-and-plating-attempt-7-samples-3414-18-sept-2-colonies-in-sample-3-none-for-4-and-14-21-sept-all-strains-13-variants-from-randomized-sample-3-11-model-generated-sequences-streaked-on-agar-plates-22-sept-colonies-picked-and-overnight-1ml-cultures-made-and-incubated-for-14-hrs-23-sept-fluorescence-assay-first-batch-of-results-obtained-25-sept-colonies-picked-from-streaked-plates-and-cultured-overnight-26-sept-plasmid-isolation-qiagen-kit-protocol-plasmid-concentration-obtained-were-very-poor-b3N-sub-37"
>
Colonies picked from streaked plates and cultured overnight.
<br/>
<br/>
</span>
<span
class=
"sept-transformation-plating-electroporation-was-done-for-all-the-samples-in-l-lactis-competent-cells-samples-were-incubated-for-1hr-after-which-they-were-spread-plated-and-incubated-for-24-hours-3-sept-colonies-in-all-plates-except-samples-4-and-14-10-colonies-were-observed-in-plate-3-colonies-were-picked-and-cultured-overnight-2-cultures-from-each-non-randomized-sample-all-10-cultures-of-each-colony-from-the-randomized-sample-3-4-sept-glycerol-added-and-strains-stored-in-80c-5-sept-transformation-and-plating-attempt-2-samples-3414-working-to-obtain-more-strains-of-randomized-sample-3-6-sept-no-colonies-observed-8-sept-transformation-and-plating-attempt-4-samples-3414-9-sept-no-colonies-observed-11-sept-transformation-and-plating-attempt-5-samples-3414-slightly-reduced-antibiotic-concentration-12-sept-golden-gate-assembly-set-up-from-left-over-plasmid-for-3414-14-sept-transformation-and-plating-attempt-6-samples-3414-15-sept-1-colony-for-sample-3-none-for-4-and-14-17-sept-transformation-and-plating-attempt-7-samples-3414-18-sept-2-colonies-in-sample-3-none-for-4-and-14-21-sept-all-strains-13-variants-from-randomized-sample-3-11-model-generated-sequences-streaked-on-agar-plates-22-sept-colonies-picked-and-overnight-1ml-cultures-made-and-incubated-for-14-hrs-23-sept-fluorescence-assay-first-batch-of-results-obtained-25-sept-colonies-picked-from-streaked-plates-and-cultured-overnight-26-sept-plasmid-isolation-qiagen-kit-protocol-plasmid-concentration-obtained-were-very-poor-b3N-sub-38"
>
26 Sept
<br/>
</span>
<span
class=
"sept-transformation-plating-electroporation-was-done-for-all-the-samples-in-l-lactis-competent-cells-samples-were-incubated-for-1hr-after-which-they-were-spread-plated-and-incubated-for-24-hours-3-sept-colonies-in-all-plates-except-samples-4-and-14-10-colonies-were-observed-in-plate-3-colonies-were-picked-and-cultured-overnight-2-cultures-from-each-non-randomized-sample-all-10-cultures-of-each-colony-from-the-randomized-sample-3-4-sept-glycerol-added-and-strains-stored-in-80c-5-sept-transformation-and-plating-attempt-2-samples-3414-working-to-obtain-more-strains-of-randomized-sample-3-6-sept-no-colonies-observed-8-sept-transformation-and-plating-attempt-4-samples-3414-9-sept-no-colonies-observed-11-sept-transformation-and-plating-attempt-5-samples-3414-slightly-reduced-antibiotic-concentration-12-sept-golden-gate-assembly-set-up-from-left-over-plasmid-for-3414-14-sept-transformation-and-plating-attempt-6-samples-3414-15-sept-1-colony-for-sample-3-none-for-4-and-14-17-sept-transformation-and-plating-attempt-7-samples-3414-18-sept-2-colonies-in-sample-3-none-for-4-and-14-21-sept-all-strains-13-variants-from-randomized-sample-3-11-model-generated-sequences-streaked-on-agar-plates-22-sept-colonies-picked-and-overnight-1ml-cultures-made-and-incubated-for-14-hrs-23-sept-fluorescence-assay-first-batch-of-results-obtained-25-sept-colonies-picked-from-streaked-plates-and-cultured-overnight-26-sept-plasmid-isolation-qiagen-kit-protocol-plasmid-concentration-obtained-were-very-poor-b3N-sub-39"
>
Plasmid isolation (Qiagen Kit protocol)
<br/>
Plasmid concentration obtained were very poor.
<br/>
</span>
</p>
</div>
<div
class=
"group-43-67A"
id=
"400:51"
>
<p
class=
"october-Dha"
id=
"400:53"
>
October
</p>
<p
class=
"oct-all-strained-streaked-from-glycerol-stocks-3-oct-colonies-were-picked-from-streaked-plates-and-cultured-overnight-4-oct-plasmid-isolation-qiagen-kit-protocol-5-oct-samples-sent-for-sequencing-jfv"
id=
"400:54"
>
<span
class=
"oct-all-strained-streaked-from-glycerol-stocks-3-oct-colonies-were-picked-from-streaked-plates-and-cultured-overnight-4-oct-plasmid-isolation-qiagen-kit-protocol-5-oct-samples-sent-for-sequencing-jfv-sub-0"
>
2 Oct
<br/>
</span>
<span
class=
"oct-all-strained-streaked-from-glycerol-stocks-3-oct-colonies-were-picked-from-streaked-plates-and-cultured-overnight-4-oct-plasmid-isolation-qiagen-kit-protocol-5-oct-samples-sent-for-sequencing-jfv-sub-1"
>
All strained streaked from glycerol stocks.
<br/>
<br/>
</span>
<span
class=
"oct-all-strained-streaked-from-glycerol-stocks-3-oct-colonies-were-picked-from-streaked-plates-and-cultured-overnight-4-oct-plasmid-isolation-qiagen-kit-protocol-5-oct-samples-sent-for-sequencing-jfv-sub-2"
>
3 Oct
<br/>
</span>
<span
class=
"oct-all-strained-streaked-from-glycerol-stocks-3-oct-colonies-were-picked-from-streaked-plates-and-cultured-overnight-4-oct-plasmid-isolation-qiagen-kit-protocol-5-oct-samples-sent-for-sequencing-jfv-sub-3"
>
Colonies were picked from streaked plates and cultured overnight.
<br/>
<br/>
</span>
<span
class=
"oct-all-strained-streaked-from-glycerol-stocks-3-oct-colonies-were-picked-from-streaked-plates-and-cultured-overnight-4-oct-plasmid-isolation-qiagen-kit-protocol-5-oct-samples-sent-for-sequencing-jfv-sub-4"
>
4 Oct
<br/>
</span>
<span
class=
"oct-all-strained-streaked-from-glycerol-stocks-3-oct-colonies-were-picked-from-streaked-plates-and-cultured-overnight-4-oct-plasmid-isolation-qiagen-kit-protocol-5-oct-samples-sent-for-sequencing-jfv-sub-5"
>
Plasmid isolation (Qiagen Kit protocol).
<br/>
</span>
<span
class=
"oct-all-strained-streaked-from-glycerol-stocks-3-oct-colonies-were-picked-from-streaked-plates-and-cultured-overnight-4-oct-plasmid-isolation-qiagen-kit-protocol-5-oct-samples-sent-for-sequencing-jfv-sub-6"
>
<br/>
5 Oct
<br/>
</span>
<span
class=
"oct-all-strained-streaked-from-glycerol-stocks-3-oct-colonies-were-picked-from-streaked-plates-and-cultured-overnight-4-oct-plasmid-isolation-qiagen-kit-protocol-5-oct-samples-sent-for-sequencing-jfv-sub-7"
>
Samples sent for sequencing.
<br/>
</span>
</p>
</div>
</div>
<div
style=
"
width: 1079px;
height: 431px;
left: 181px;
top: 5171px;
position: absolute;
"
>
<div
style=
"
width: 357px;
height: 156px;
left: 0px;
top: 0px;
position: absolute;
color: #221023;
font-size: 36px;
font-family: Montserrat;
font-weight: 700;
word-wrap: break-word;
"
>
October
</div>
<div
style=
"width: 1079px; left: 0px; top: 72px; position: absolute"
>
<span
style=
"
color: black;
font-size: 24px;
font-family: Montserrat;
font-weight: 700;
word-wrap: break-word;
"
>
2 Oct
<br
/></span
><span
style=
"
color: black;
font-size: 24px;
font-family: Montserrat;
font-weight: 400;
word-wrap: break-word;
"
>
All strained streaked from glycerol stocks.
<br
/></span
><span
style=
"
color: black;
font-size: 24px;
font-family: Montserrat;
font-weight: 400;
word-wrap: break-word;
"
><br
/></span
><span
style=
"
color: black;
font-size: 24px;
font-family: Montserrat;
font-weight: 700;
word-wrap: break-word;
"
>
3 Oct
<br
/></span
><span
style=
"
color: black;
font-size: 24px;
font-family: Montserrat;
font-weight: 400;
word-wrap: break-word;
"
>
Colonies were picked from streaked plates and cultured
overnight.
<br
/></span
><span
style=
"
color: black;
font-size: 24px;
font-family: Montserrat;
font-weight: 400;
word-wrap: break-word;
"
><br
/></span
><span
style=
"
color: black;
font-size: 24px;
font-family: Montserrat;
font-weight: 700;
word-wrap: break-word;
"
>
4 Oct
<br
/></span
><span
style=
"
color: black;
font-size: 24px;
font-family: Montserrat;
font-weight: 400;
word-wrap: break-word;
"
>
Plasmid isolation (Qiagen Kit protocol).
<br
/></span
><span
style=
"
color: black;
font-size: 24px;
font-family: Montserrat;
font-weight: 700;
word-wrap: break-word;
"
><br
/>
5 Oct
<br
/></span
><span
style=
"
color: black;
font-size: 24px;
font-family: Montserrat;
font-weight: 400;
word-wrap: break-word;
"
>
Samples sent for sequencing.
<br
/></span>
</div>
</div>
<img
style=
"
width: 907px;
height: 793px;
left: 267px;
top: 687px;
position: absolute;
"
src=
"https://static.igem.wiki/teams/4931/wiki/wetlab.png"
/>
{% endblock %}
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