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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: Stability Constructions</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 10 ======= -->
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| - | <div class="panel panel-info notes-panel">
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| - | <span class="note-tag" id="week10"></span>
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| - | <div class="panel-heading">
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| - | <h3 class="panel-title">Week 10 : 09/01/2014 ➡ 08/07/2014 <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="#0903" role="tab" data-toggle="tab" class="date-notebook">09/03/14</a></li>
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| - | <li><a href="#0904" role="tab" data-toggle="tab" class="date-notebook">09/04/14</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="0903">
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| - | <ul class="list-notebook">
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| - | <li>
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| - | <p>Construction of stability epitopes : the main idea is to use the primers 57-58 and 59 as matrices in order to synthesize and amplify the stability epitope. The PCR uses the iGEM PCR protocol with Q5 polymerase. The temperature of annealing is 55°C, the elongation time is 5", the number of cycle is 25.</p>
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| - | <p>
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| - | Mix:
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| - | <ul>
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| - | <li>5µL OiGEM-57 or 58 or 59</li>
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| - | <li>2,5µL OiGEM-60</li>
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| - | <li>2,5µL OiGEM-44</li>
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| - | <li>25µL Q5 2x Mix</li>
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| - | <li>15µL H2O</li>
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| - | </ul>
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| - | </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/a/a0/AMU_Team-stabs-0903.png" style="height:350px">
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| - | </div>
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| - | </li>
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| - | <li>
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| - | <p>Clean-up of these PCR and elution with 30µL H2O at 70°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="0904">
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| - | <ul class="list-notebook">
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| - | <li>
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| - | <p>Digestion ES of PCR products of the stability epitopes and of piGEM-01.13 (pSB1C3-BBa_J04450).</p>
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| - | <p>
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| - | Mix:
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| - | <ul>
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| - | <li>10µL K10512-06 or 07 or 08 or 5µL piGEM-01.13</li>
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| - | <li>1µL EcoRI</li>
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| - | <li>1µL SpeI</li>
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| - | <li>5µL Buffer 2.1</li>
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| - | <li>qsp 50µL of water</li>
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| - | </ul>
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| - | </p><br>
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| - | <p>Incubation 60' at 37°C.</p>
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| - | </li>
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| - | <li>
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| - | <p>Then, for K10512-06 or 07 or 08, inactivation of enzymes 20' at 80°C.</p>
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| - | </li>
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| - | <li>
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| - | <p>Clean-up of piGEM-01.13 and elution with 30µL of water. Add 1µL of SAP (Promega) + 2µL of SAP buffer (dephosphorylation). Incubation 30' at 37°C, then Inactivation of enzyme 20' at 65°C.</p>
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| - | </li>
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| - | <li>
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| - | <p>Ligation K10512-06 or 07 or 08 with piGEM-01.13.</p>
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| - | <p>
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| - | Mix:
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| - | <ul>
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| - | <li>2µL piGEM-01.13</li>
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| - | <li>4µL K10512-06 or 07 or 08</li>
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| - | <li>1µL T4 ligase 10U/µL</li>
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| - | <li>2µL T4 ligase buffer</li>
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| - | <li>11µL 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>Transformation of 90µL of E.coli TG1 competent cells with 10µL of ligation. The selective medium is LB-Cm 30µg/mL.</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>
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| - | </div>
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| - | </div>
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| - |
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| - | <!-- ======= WEEK 11 ======= -->
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| - | <div class="panel panel-info notes-panel last-note">
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| - | <span class="note-tag" id="week11"></span>
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| - | <div class="panel-heading">
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| - | <h3 class="panel-title">Week 11 : 09/08/2014 ➡ 08/14/2014 <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="#0908" role="tab" data-toggle="tab" class="date-notebook">09/08/14</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="0908">
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| - | <ul class="list-notebook">
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| - | <li>
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| - | <p>Test 5 clones of each transformation (with pSB1C3- K10512-06 or 07 or 08) by PCR. The temperature of annealing is 55°C, the elongation time is 1'</p>
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| - | <p>
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| - | Mix:
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| - | <ul>
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| - | <li>2µL OiGEM-60</li>
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| - | <li>2µL OiGEM-44</li>
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| - | <li>4µL dNTP</li>
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| - | <li>1µL DMSO</li>
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| - | <li>2µL buffer</li>
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| - | <li>0,5µL Taq polymérase</li>
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| - | <li>8,5µL H2O</li>
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| - | </ul>
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| - | </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/2014/0/02/AMU_Team-stabs-0908.png" style="width:350px">
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| - | <div class="media-body" style="padding-left: 10px;">
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| - | <p>All clones are good except the K10512-08-2. The pSB1C3-K10512-06-1, pSB1C3-K10512-07-1 and pSB1C3-K10512-08-1 are sent to sequencing. They are 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="#week10" class="list-group-item">
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| - | <h4 class="list-group-item-heading">Week 10
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| - | <small style="padding-left:20px">09/01/14 → 09/07/14</small>
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| - | </h4>
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| - | </a>
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| - | <a data-scroll href="#week11" class="list-group-item">
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| - | <h4 class="list-group-item-heading">Week 11
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| - | <small style="padding-left:20px">09/08/14 → 09/14/14</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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