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Commit 42c7df62 authored by Devmc's avatar Devmc
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notebook

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static/utils.css
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width: 75% !important;
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width: 65% !important;
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wiki/pages/notebook.html
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<h1 class="content-header2">Notebook</h1>
<section>
<h2></h2>
<p></p>
<h2>7/1</h2>
<p>Team ice-breaking.</p>
<p>Team discussion（Team name, uniform, logo...）.</p>
<div class="imager">
<img class="rw-65" src="https://static.igem.wiki/teams/4281/wiki/notebook/t-ecnuas-notebook-01.jpg" alt="">
</div>
</section>
<section>
<h2>7/2</h2>
<p>Laboratory safety training.</p>
<p>Configuring the culture medium.</p>
<p>Inoculation of the <i>Streptomyces rapamycinicus</i>, 30℃.</p>
<p>Inoculation of the strain containing plasmid pKC1139.</p>
<div class="imager">
<img class="rw-65" src="https://static.igem.wiki/teams/4281/wiki/notebook/t-ecnuas-notebook-02.jpg" alt="">
</div>
</section>
<section>
<h2>7/3</h2>
<p>Design the questionnaires.</p>
<p>Extract plasmid pKC1139 using a plasmid extract kit.</p>
<div class="imager">
<img class="rw-65" src="https://static.igem.wiki/teams/4281/wiki/notebook/t-ecnuas-notebook-03.jpg" alt="">
</div>
</section>
<section>
<h2>7/4</h2>
<p>Team collaboration.</p>
<div class="imager">
<img class="rw-65" src="https://static.igem.wiki/teams/4281/wiki/notebook/t-ecnuas-notebook-04.jpg" alt="">
</div>
<br>
<p>Amplify target DNA fragments by PCR. </p>
<br>
<p>PCR result:</p>
<p>Line 1, 3, and 5 are upstream homologous gene fragments, and line 2, 4, and 6 are downstream homologous gene
fragments.</p>
<div class="imager">
<img class="rw-65" src="https://static.igem.wiki/teams/4281/wiki/notebook/t-ecnuas-notebook-05.jpg" alt="">
</div>
<br>
<p>Line 1, 4 and 7 are plasmid pKC1139.</p>
<p>Line2,3,5,6,8,9,10 are double enzyme digested plasmid pKC1139 (EcoRI/HindIII).</p>
<p>Construct recombinant plasmids by Homologous reorganization.</p>
</section>
<section>
<h2>7/5</h2>
<p>Verification of the recombinant plasmids pKC-M271_ 14685/ M271_ 14690through colony PCR.</p>
<p>Inoculation of the strain containing correct plasmids.</p>
</section>
<section>
<h2>7/6</h2>
<p>Extract the plasmids pKC-M271_ 14685/ M271_ 14690 with a plasmids extraction kit.</p>
<p>Double enzyme digestion verification.</p>
<div class="imager">
<img class="rw-65" src="https://static.igem.wiki/teams/4281/wiki/notebook/t-ecnuas-notebook-06.jpg" alt="">
</div>
<br>
<p>Line 1,3,5 are recombinant plasmid pKC-M271_ 14685/ M271_ 14690.</p>
<p>Line 2,4,6 are recombinant plasmid pKC-M271_14685/ M271_14690 digested with EcoR I/Hind III</p>
</section>
<section>
<h2>7/7</h2>
<p>Expert interview.</p>
<p>Transfer plasmids pKC-M271_ 14685/ M271_ 14690 into ET12567/pUZ8002 competent cells, and coat to LB plate
containing antibodies, 37℃ incubate overnight.</p>
<div class="imager">
<img class="rw-65" src="https://static.igem.wiki/teams/4281/wiki/notebook/t-ecnuas-notebook-07.jpg" alt="">
</div>
</section>
<section>
<h2>7/8</h2>
<p>Take fixed makeup photos.</p>
<p>Pick up single colonies and inoculate them in 4ml fresh culture medium,37℃ 220rpm overnight.</p>
</section>
<section>
<h2>7/9</h2>
<p>
1. Transfer 1ml strain suspension into 50ml fresh culture medium, when OD600 is around 0.4-0.6, centrifuge
and discard the medium, wash it twice with LB (no antibody contained) and resuspended it with 500ul
culture medium.
</p>
<p>
2. Collect the <i>Streptomyces rapamycinicus</i> spore through 2×YT culture medium which was incubated on the
plate.
</p>
<p>
3. Mix the <i>E. coli</i> suspension from step (1) and the spore suspension from step (2), coat onto M-ISP4
solid medium, and incubate at 30°C.
</p>
</section>
<section>
<h2>7/10</h2>
<p>After incubated on the M-ISP4 solid medium for 16h, add antibodies, and incubate at 30℃ for 7 days.</p>
</section>
<section>
<h2>7/17</h2>
<p>PCR identification of colony.</p>
<p>Screening for single cross-over strains.</p>
<p>Filtrate single cross-over strains.</p>
<div class="imager">
<img class="rw-65" src="https://static.igem.wiki/teams/4281/wiki/notebook/t-ecnuas-notebook-08.jpg" alt="">
</div>
</section>
<section>
<h2>7/23</h2>
<p>Carry out educational activities.</p>
<p>Expert interview.</p>
<p>Screening double cross-over strains.</p>
<p>The correct strain was streaked on the oat solid medium and cultured at 30°C for 7 days</p>
<div class="imager">
<img class="rw-65" src="https://static.igem.wiki/teams/4281/wiki/notebook/t-ecnuas-notebook-09.jpg" alt="">
</div>
</section>
<section>
<h2>7/29</h2>
<p>Expert interview.</p>
<p>Writing implementation.</p>
</section>
<section>
<h2>7/30</h2>
<p>Fermentation culture of <i>Streptomyces rapamycinicus</i>. </p>
</section>
<section>
<h2>8/6</h2>
<p>Pick up colonies and inoculate them in 3ml fresh fermentation culture medium,28℃ 220rpm overnight.</p>
</section>
<section>
<h2>8/7-8/18</h2>
<p>Transfer 3ml strain suspension into 30ml fresh fermentation culture medium, culture at 28℃ 200rpm, and
collect samples at 5d, 7d, 9d, and 11d. </p>
<p>Rapamycin production estimation by HPLC.</p>
<div class="imager">
<img class="rw-65" src="https://static.igem.wiki/teams/4281/wiki/notebook/t-ecnuas-notebook-10.jpg" alt="">
</div>
</section>
<section>
<h2>8/19</h2>
<p>Write wiki.</p>
<p>Make a presentation video.</p>
<p>Prepare for presentation.</p>
<div class="imager">
<img class="rw-65" src="https://static.igem.wiki/teams/4281/wiki/notebook/t-ecnuas-notebook-11.jpg" alt="">
</div>
</section>
</div>
......
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