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| - | {{Team:Aix-Marseille/header}}
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| - | <html>
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| - | <script>
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| - | // Function to handle the movement of the sidebar in the Notebook
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| - | $(document).ready(function () {
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| - | var sidebar = $('#sidebar-notebook');
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| - | var sidebarContainer = $('#sidebar-notebook-div');
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| | | | |
| - | $(window).scroll(function (event) {
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| - | var ypos = $(this).scrollTop();
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| - | var scrollTop = $(window).scrollTop(),
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| - | elementOffset = sidebar.offset().top,
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| - | distance = (elementOffset - scrollTop);
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| - |
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| - | var collision_top = sidebar.overlaps($('#stop_top'));
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| - | var collision_bot = sidebar.overlaps($('#stop_bot'));
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| - |
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| - | if (collision_top.hits.length) { /* Collision TOP */
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| - | // To fix at the top of the .row div
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| - | sidebar.css({ top: '',
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| - | bottom: '',
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| - | position: '',
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| - | width: ''
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| - | });
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| - | sidebarContainer.css({ position:'',
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| - | bottom: '',
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| - | right: ''
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| - | });
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| - | if (distance < 141) {
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| - | // To fix at the top of the page while scrolling
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| - | sidebar.css({ top: '141px',
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| - | bottom: '',
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| - | position: 'fixed',
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| - | width: '262.5px'
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| - | });
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| - | sidebarContainer.css({ position: '',
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| - | bottom: '',
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| - | right: ''
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| - | });
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| - | }
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| - | }
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| - | else if (collision_bot.hits.length) { /* Collision BOTTOM */
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| - | // To fix at the bottom of the .row div
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| - | sidebar.css({ bottom: '',
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| - | top: '',
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| - | position: '',
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| - | width: ''
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| - | });
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| - | sidebarContainer.css({ position: 'absolute',
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| - | bottom: '0px',
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| - | right: '0px'
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| - | });
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| - | if (distance > 141) {
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| - | // To fix at the top of the page while scrolling
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| - | sidebar.css({ top: '141px',
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| - | bottom: '',
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| - | position: 'fixed',
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| - | width: '262.5px'
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| - | });
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| - | sidebarContainer.css({ position: '',
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| - | bottom: '',
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| - | right: ''
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| - | });
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| - | }
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| - | }
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| - | else {
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| - | // To fix at the top of the page while scrolling
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| - | sidebar.css({ top: '141px',
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| - | bottom: '',
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| - | position: 'fixed',
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| - | width: '262.5px'
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| - | });
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| - | sidebarContainer.css({ position: '',
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| - | bottom: '',
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| - | right: ''
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| - | });
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| - | }
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| - | });
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| - | });
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| - | </script>
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| - |
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| - |
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| - |
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| - | <div class="container">
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| - | <h1 class="project-title">Notebook: SerA</h1>
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| - | <br><br>
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| - | <div id="stop_top"></div>
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| - | <div class="row" style="position:relative;" id="notebook-row">
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| - | <div class="col-md-9" id="notes"><!-- NOTES -->
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| - |
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| - | <!-- ======= WEEK 9 ======= -->
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| - | <div class="panel panel-info notes-panel last-note">
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| - | <span class="note-tag" id="week9"></span>
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| - | <div class="panel-heading">
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| - | <h3 class="panel-title">Week 9 : 08/25/2014 ➡ 08/31/2014 <div class="pull-right"></div></h3>
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| - | </div>
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| - | <div class="panel-body">
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| - |
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| - | <ul class="nav nav-tabs" role="tablist">
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| - | <li class="active"><a href="#0826" role="tab" data-toggle="tab" class="date-notebook">08/26/14</a></li>
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| - | <li><a href="#0827" role="tab" data-toggle="tab" class="date-notebook">08/27/14</a></li>
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| - | <li><a href="#0828" role="tab" data-toggle="tab" class="date-notebook">08/28/14</a></li>
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| - | </ul>
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| - |
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| - | <div class="tab-content">
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| - |
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| - | <div class="tab-pane fade in active" id="0826">
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| - | <ul class="list-notebook">
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| - | <li>
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| - | <p>Clean-up of PCR product number 82 and 83 from 8-8-14 (50µL)</p>
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| - | </li>
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| - | <li>
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| - | <p>SOE reaction (= overlap PCR):</p>
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| - | <p>
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| - | Mix:
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| - | <ul>
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| - | <li>PCR product 82= 3µL</li>
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| - | <li>PCR product 83 = 3µL</li>
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| - | <li>dNTP 10mM = 2µL</li>
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| - | <li>Q5 buffer = 8µL</li>
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| - | <li>Q5 polymerase = 0,5µL</li>
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| - | <li>H2O qsp 40µL</li>
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| - | </ul>
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| - | </p><br>
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| - | <p>
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| - | Program:
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| - | <ol>
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| - | <li>98°C 2’</li>
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| - | <li>98°C 20”</li>
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| - | <li>45°C 25”</li>
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| - | <li>72°C 1’</li>
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| - | <li>72°C 10’</li>
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| - | </ol>
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| - | Repeat the steps 3-4-5, 10 times.
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| - | </p>
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| - | </li>
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| - | </ul>
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| - | </div>
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| - |
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| - | <div class="tab-pane fade in" id="0827">
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| - | <ul class="list-notebook">
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| - | <li>
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| - | <p>PCR2: Second PCR round to amplification of serA from the SOE product (26-08-14) with primers 5 and 6, using the iGEM PCR protocol with Q5 polymerase. The temperature of annealing is 60°C</p>
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| - | <div class="media notes-media">
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| - | <img class="media-object img-rounded pull-right" src="https://static.igem.org/mediawiki/2014/8/80/AMU_Team-serA-0827.png" style="height:350px">
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| - | <div class="media-body" style="padding-left: 10px;">
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| - | <p>Result: the amplification of serA is good. Clean-up of serA (PCR2) and elution with 45µL of water.</p>
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| - | </div>
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| - | </div>
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| - | </li>
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| - | <li>
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| - | <p>Digestion of pSB1A3 and pSB1C3 during 1h30 at 37°C. Then, inactivation of enzymes during 20’ at 80°C</p>
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| - | <p>
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| - | Mix:
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| - | <ul>
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| - | <li>10µL of plasmid</li>
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| - | <li>1µL EcoRI</li>
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| - | <li>1µL PstI</li>
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| - | <li>5µL Buffer 2.1</li>
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| - | <li>33 µL H2O</li>
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| - | </ul>
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| - | </p>
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| - | </li>
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| - | <li>
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| - | <p>Digestion of the insert (serA) during 1h at 37°C. Then inactivation of enzymes during 20’ at 80°C</p>
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| - | <p>
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| - | Mix:
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| - | <ul>
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| - | <li>20µL of serA</li>
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| - | <li>1µL EcoRI</li>
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| - | <li>1µL PstI</li>
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| - | <li>5µL Buffer 2.1</li>
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| - | <li>13 µL H2O</li>
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| - | </ul>
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| - | </p>
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| - | </li>
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| - | <li>
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| - | <p>NEB ligation:</p>
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| - | <p>
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| - | <ul>
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| - | <li>2µL of digested plasmid </li>
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| - | <li>2µL of digested serA insert</li>
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| - | <li>2µL ligase buffer</li>
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| - | <li>1µL ligase (NEB)</li>
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| - | <li>13 µL H2O</li>
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| - | </ul>
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| - | </p><br>
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| - | <p>Incubation overnight at 16°C</p>
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| - | </li>
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| - | </ul>
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| - | </div>
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| - |
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| - | <div class="tab-pane fade in" id="0828">
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| - | <ul class="list-notebook">
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| - | <li>
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| - | <p>Transformation of 90µL of DH5α super competent cells with 10µL of ligation (pSB1C3-SerA or pSB1A2-SerA). The selective medium is LB + Ampicillin (Ap) 100µg/mL or LB + Chloramphenicol (Cm) 30µg/mL</p>
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| - | <div class="media notes-media">
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| - | <img class="media-object img-rounded pull-right" src="https://static.igem.org/mediawiki/2014/6/6e/AMU_Team-serA-0828.png" style="height:350px">
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| - | <div class="media-body" style="padding-left: 10px;">
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| - | <p>pSB1C3-SerA-1 seemed good. It was sent to sequencing. It’s OK</p>
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| - | </div>
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| - | </div>
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| - | </li>
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| - | </ul>
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| - | </div>
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| - |
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| - | </div>
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| - | </div>
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| - | </div>
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| - |
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| - | </div><!-- /NOTES -->
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| - |
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| - | <!-- SIDEBAR -->
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| - | <div class="col-md-3" id="sidebar-notebook-div">
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| - | <div class="list-group" id="sidebar-notebook">
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| - | <a data-scroll href="#week9" class="list-group-item">
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| - | <h4 class="list-group-item-heading">Week 9
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| - | <small style="padding-left:20px">08/25/14 → 08/31/14</small>
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| - | </h4>
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| - | </a>
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| - |
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| - | </div>
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| - | </div>
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| - | </div>
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| - | <div id="stop_bot"></div>
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| - | </div>
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| - | </html>
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| - |
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| - | {{Team:Aix-Marseille/footer}}
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