From 27b9f4f293d05fd67d6b3597004efd11add364a8 Mon Sep 17 00:00:00 2001
From: anvin-f <anvinfu@gmail.com>
Date: Wed, 2 Oct 2024 08:57:19 +0800
Subject: [PATCH] e
---
wiki/pages/engineering.html | 6 ++---
wiki/pages/protocols.html | 48 ++++++++++++++++++-------------------
2 files changed, 27 insertions(+), 27 deletions(-)
diff --git a/wiki/pages/engineering.html b/wiki/pages/engineering.html
index f29a95c..f02b883 100644
--- a/wiki/pages/engineering.html
+++ b/wiki/pages/engineering.html
@@ -181,7 +181,7 @@ endblock %}
<p><span>Aim: to build a plasmid (pENTR1A-PSMA-GFP) that allows us to detect PSMA-positive prostate cancer cells by measuring the fluorescence by GFP.</span></p>
<br>
-
+ <a name = "build1"></a>
<h5><span>Build</span></h5>
<p><span>We purchased the pENTR1A backbone and another plasmid containing the PSMA promoter and GFP gene. Both plasmids were digested and gel electrophoresis was performed.</span></p>
<p><span>We then purified the agarose gel and the two genes were ligated to form plasmid pENTR1A-PSMA-GFP. (for more information refer to</span><a href = "#cycle3"> cycle 3</a><span>)</span></p>
@@ -189,7 +189,7 @@ endblock %}
<p><span>After the ligated plasmids were extracted, we transfected different concentrations of them into multiple concentrations of PSMA-positive cancer cell line MLLB-2.</span></p>
<br>
-
+ <a name = "test1"></a>
<h5><span>Test</span></h5>
<p><span>Fluorescence given out by GFP was visualised by using an inverted microscope with a live cell-monitoring system.</span></p>
@@ -339,7 +339,7 @@ endblock %}
<br>
- <a name = "desgin4"></a>
+ <a name = "design4"></a>
<h5><span>Design</span></h5>
<p><span>We followed a set of standard protocols, found in manufacturers’ product sheets, during our process of plasmid construction.</span></p>
diff --git a/wiki/pages/protocols.html b/wiki/pages/protocols.html
index c74ac0e..7d78eb6 100644
--- a/wiki/pages/protocols.html
+++ b/wiki/pages/protocols.html
@@ -700,8 +700,8 @@
<h5>Procedure</h5>
<div class="border1">
<ol>
- <li style="font-weight: 400;"><strong>Add </strong><span style="font-weight: 400;">20 μL/well of </span><strong>medium</strong><span style="font-weight: 400;"> from the transfected cells </span><strong>within</strong> <strong>72 hours</strong><span style="font-weight: 400;"> after transfection</span> <span style="font-weight: 400;">to a black opaque 96-well plate. (For Transfection Protocol, please refer 3.4 Transfection)</span></li>
- <li><span style="font-weight: 400;">Add 50 μL of </span><strong>coelenterazine working solution</strong><span style="font-weight: 400;"> to each well and </span><strong>detect the light output </strong><span style="font-weight: 400;">immediately</span></li>
+ <li ><strong>Add </strong><span >20 μL/well of </span><strong>medium</strong><span > from the transfected cells </span><strong>within</strong> <strong>72 hours</strong><span > after transfection</span> <span >to a black opaque 96-well plate. (For Transfection Protocol, please refer 3.4 Transfection)</span></li>
+ <li><span >Add 50 μL of </span><strong>coelenterazine working solution</strong><span > to each well and </span><strong>detect the light output </strong><span >immediately</span></li>
</ol>
</div>
</div>
@@ -709,8 +709,8 @@
<div class="border2">
<h5>Materials</h5>
<ul>
- <li style="font-weight: 400;"><span style="font-weight: 400;">Pierce™ Gaussia Luciferase Flash Assay Kit</span></li>
- <li style="font-weight: 400;"><span style="font-weight: 400;">Black Opaque 96-well Plate</span></li>
+ <li ><span >Pierce™ Gaussia Luciferase Flash Assay Kit</span></li>
+ <li ><span >Black Opaque 96-well Plate</span></li>
</ul>
</div>
</div>
@@ -728,13 +728,13 @@
<div class="border1">
<ol>
- <li style="font-weight: 400;"><span style="font-weight: 400;">Remove original culture medium and add 100 µL of fresh medium</span></li>
- <li style="font-weight: 400;"><span style="font-weight: 400;">Add 10 µL of the 12-mM MTT solution to each well. Include a negative control by adding 10 µL of the MTT solution to 100 µL of medium alone.</span></li>
- <li style="font-weight: 400;"><span style="font-weight: 400;">Incubate at 37°C for 2–5 hours. </span></li>
- <li style="font-weight: 400;"><span style="font-weight: 400;">Remove 85 µL of medium from the wells.</span></li>
- <li style="font-weight: 400;"><span style="font-weight: 400;">Add 50 µL of DMSO to each well, then pipet up and down thoroughly to mix.</span></li>
- <li style="font-weight: 400;"><span style="font-weight: 400;">Incubate at 37°C for 10 minutes. </span></li>
- <li style="font-weight: 400;"><span style="font-weight: 400;">Pipet up and down to mix each sample again, then read the absorbance at 540 nm. </span></li>
+ <li ><span >Remove original culture medium and add 100 µL of fresh medium</span></li>
+ <li ><span >Add 10 µL of the 12-mM MTT solution to each well. Include a negative control by adding 10 µL of the MTT solution to 100 µL of medium alone.</span></li>
+ <li ><span >Incubate at 37°C for 2–5 hours.</span></li>
+ <li ><span >Remove 85 µL of medium from the wells.</span></li>
+ <li ><span >Add 50 µL of DMSO to each well, then pipet up and down thoroughly to mix.</span></li>
+ <li ><span >Incubate at 37°C for 10 minutes.</span></li>
+ <li ><span >Pipet up and down to mix each sample again, then read the absorbance at 540 nm.</span></li>
</ol>
@@ -745,10 +745,10 @@
<h5>Materials</h5>
<ul>
- <li style="font-weight: 400;"><span style="font-weight: 400;">MTT Cell Viability Assay Kit</span></li>
- <li style="font-weight: 400;"><span style="font-weight: 400;">PBS</span></li>
- <li style="font-weight: 400;"><span style="font-weight: 400;">DMSO</span></li>
- <li style="font-weight: 400;"><span style="font-weight: 400;">Complete culture medium</span></li>
+ <li ><span >MTT Cell Viability Assay Kit</span></li>
+ <li ><span >PBS</span></li>
+ <li ><span >DMSO</span></li>
+ <li ><span >Complete culture medium</span></li>
</ul>
</div>
</div>
@@ -896,15 +896,15 @@
<div class="border1">
<h6>Procedure</h6>
<ol>
- <li style="font-weight: 400;"><strong>Dilute</strong><span style="font-weight: 400;"> the poly-D-lysine solution with </span><strong>sterile DPBS</strong><span style="font-weight: 400;"> to prepare a 50 μg/mL working solution</span></li>
- <li style="font-weight: 400;"><strong>Coat</strong><span style="font-weight: 400;"> the surface of 75 cm</span><span style="font-weight: 400;">2</span><span style="font-weight: 400;"> culture vessel with 11.7 mL </span><strong>working solution</strong><span style="font-weight: 400;"> of poly-D-lysine</span></li>
- <li style="font-weight: 400;"><strong>Incubate</strong><span style="font-weight: 400;"> the vessel at </span><strong>room temperature</strong><span style="font-weight: 400;"> for an hour then </span><strong>remove</strong><span style="font-weight: 400;"> the poly-D-lysine solution with a pipette gun.</span></li>
- <li style="font-weight: 400;"><strong>Rinse</strong><span style="font-weight: 400;"> culture surface for 3 times with 24 mL distilled water and remove it from the flask.</span></li>
- <li style="font-weight: 400;"><span style="font-weight: 400;">Leave the coated culture vessel uncovered in the biosafety cabinet to </span><strong>dry</strong><span style="font-weight: 400;"> for </span><strong>2 hours</strong><span style="font-weight: 400;">. Afterwards, the flask can be used immediately or store at </span><strong>4°C</strong></li>
- <li style="font-weight: 400;"><strong>Dilute</strong><span style="font-weight: 400;"> the thawed laminin solution to 3 μg/mL with </span><strong>sterile distilled water</strong><span style="font-weight: 400;">.</span></li>
- <li style="font-weight: 400;"><span style="font-weight: 400;">Add </span><strong>laminin solution</strong><span style="font-weight: 400;"> into poly-D-lysine coated culture vessel to cover the whole surface.</span></li>
- <li style="font-weight: 400;"><strong>Incubate</strong><span style="font-weight: 400;"> and store in a </span><strong>37°C 5% CO</strong><strong>2</strong><strong> incubator</strong><span style="font-weight: 400;">.</span></li>
- <li><strong> Aspirate</strong><span style="font-weight: 400;"> laminin solution from coated culture vessel and let it </span><strong>dry</strong><span style="font-weight: 400;"> right before seeding cells.</span></li>
+ <li ><strong>Dilute</strong><span > the poly-D-lysine solution with </span><strong>sterile DPBS</strong><span > to prepare a 50 μg/mL working solution</span></li>
+ <li ><strong>Coat</strong><span > the surface of 75 cm</span><span >2</span><span > culture vessel with 11.7 mL </span><strong>working solution</strong><span > of poly-D-lysine</span></li>
+ <li ><strong>Incubate</strong><span > the vessel at </span><strong>room temperature</strong><span > for an hour then </span><strong>remove</strong><span > the poly-D-lysine solution with a pipette gun.</span></li>
+ <li ><strong>Rinse</strong><span > culture surface for 3 times with 24 mL distilled water and remove it from the flask.</span></li>
+ <li ><span >Leave the coated culture vessel uncovered in the biosafety cabinet to </span><strong>dry</strong><span > for </span><strong>2 hours</strong><span >. Afterwards, the flask can be used immediately or store at </span><strong>4°C</strong></li>
+ <li ><strong>Dilute</strong><span > the thawed laminin solution to 3 μg/mL with </span><strong>sterile distilled water</strong><span >.</span></li>
+ <li ><span >Add </span><strong>laminin solution</strong><span > into poly-D-lysine coated culture vessel to cover the whole surface.</span></li>
+ <li ><strong>Incubate</strong><span > and store in a </span><strong>37°C 5% CO</strong><strong>2</strong><strong> incubator</strong><span >.</span></li>
+ <li><strong> Aspirate</strong><span > laminin solution from coated culture vessel and let it </span><strong>dry</strong><span > right before seeding cells.</span></li>
</ol>
</div>
--
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