Difference between revisions of "Team:NCKU Tainan/Protocol"
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<h1 class="head2">Protocol</h1> | <h1 class="head2">Protocol</h1> | ||
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| − | + | <p class="pcontent">1. Gently mix the following reaction by pipetting and centrifuge briefly.</p> | |
| − | + | <table class="card card-body"> | |
| − | + | <tr> | |
| − | + | <th></th> | |
| − | + | <th>20 μl system</th> | |
| − | + | <th>50 μl system</th> | |
| + | </tr> | ||
| − | + | <tr> | |
| − | + | <td>Template</td> | |
| − | + | <td>12～20 ng</td> | |
| − | + | <td>30～50 ng</td> | |
| − | + | </tr> | |
| − | + | <tr> | |
| − | + | <td>Forward primer</td> | |
| − | + | <td>1.0 μl</td> | |
| − | + | <td>2.5 μl</td> | |
| − | + | </tr> | |
| − | + | <tr> | |
| − | + | <td>Reverse primer</td> | |
| − | + | <td>1.0 μl</td> | |
| − | + | <td>2.5 μl</td> | |
| − | + | </tr> | |
| − | + | <tr> | |
| − | + | <td>dNTP</td> | |
| − | + | <td>1.6 μl</td> | |
| − | + | <td>4.0 μl</td> | |
| − | + | </tr> | |
| − | + | <tr> | |
| − | + | <td>10x Buffer</td> | |
| − | + | <td>2.0 μl</td> | |
| − | + | <td>5.0 μl</td> | |
| − | + | </tr> | |
| − | + | </table> | |
| + | <p class="pcontent">Program of KOD DNA polymerase</p> | ||
| + | <table class="card card-body"> | ||
| + | <tr> | ||
| + | <th>Temperature</th> | ||
| + | <th>Time</th> | ||
| + | <th></th> | ||
| + | </tr> | ||
| − | + | <tr> | |
| − | + | <td>94 ℃</td> | |
| − | + | <td>3 min.</td> | |
| − | + | <td rowspan="3">25～30 cycles</td> | |
| − | + | </tr> | |
| − | + | ||
| − | + | <tr> | |
| − | + | <td>94 ℃</br>(Denaturation)</td> | |
| − | + | <td>40 sec</td> | |
| − | + | </tr> | |
| − | + | ||
| − | + | <tr> | |
| − | + | <td>57.5 ℃</br>(Annealing)</td> | |
| − | + | <td>30 sec</td> | |
| − | + | </tr> | |
| − | + | <tr> | |
| − | + | <td>72 ℃</br>(Extension)</td> | |
| − | + | <td>Depend on sequence size</br>(2 kbp/min. for Taq)</td> | |
| − | + | <td></td> | |
| + | </tr> | ||
| − | + | <tr> | |
| − | + | <td>72 ℃</td> | |
| − | + | <td>5 min.</td> | |
| − | + | <td></td> | |
| − | + | </tr> | |
| − | + | <tr> | |
| − | + | <td>4 ℃</td> | |
| − | + | <td>∞</td> | |
| − | + | <td></td> | |
| − | + | </tr> | |
| − | + | </table> | |
| − | + | <p class="pcontent">2. Confirm the size of the digested product by gel electrophoresis.</br> | |
| − | + | 3. Gel purification of the target size.</p> | |
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| − | </ | + | |
| + | </div> | ||
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| + | </div> | ||
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| + | <div id="list-item-2"> | ||
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| + | <div> | ||
| + | <div class="row"> | ||
| + | <a class="btn col-md-12" data-toggle="collapse" href="#test" role="button" aria-expanded="false" | ||
| + | aria-controls="multiCollapseExample1"> | ||
| + | test | ||
| + | <i class="fa fa-arrow-down fa-10" aria-hidden="true"></i> | ||
| + | </a> | ||
| + | </div> | ||
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| + | <div class="collapse multi-collapse" id="test""> | ||
Revision as of 15:14, 21 September 2018