Oscarliu117 (Talk | contribs) |
Oscarliu117 (Talk | contribs) |
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<div id="sidelist" class="list-Fgroup"> | <div id="sidelist" class="list-Fgroup"> | ||
<a class="list-group-item list-group-item-action" href="#list-item-1">PCR</a> | <a class="list-group-item list-group-item-action" href="#list-item-1">PCR</a> | ||
− | <a class="list-group-item list-group-item-action" href="#list-item-2"> | + | <a class="list-group-item list-group-item-action" href="#list-item-2">Plasmid Construction</a> |
− | <a class="list-group-item list-group-item-action" href="#list-item-3"> | + | <a class="list-group-item list-group-item-action" href="#list-item-3">PCR Clean-Up & Gel Extraction</a> |
<a class="list-group-item list-group-item-action" href="#"><i class="fa fa-arrow-up fa-1x" | <a class="list-group-item list-group-item-action" href="#"><i class="fa fa-arrow-up fa-1x" | ||
aria-hidden="true"></i></a> | aria-hidden="true"></i></a> | ||
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</table> | </table> | ||
− | <p class="pcontent"></br> | + | <p class="pcontent"></br> Program of KOD DNA |
+ | polymerase</p> | ||
<table class="card card-body PCR2"> | <table class="card card-body PCR2"> | ||
<tr> | <tr> | ||
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<div class="collapse multi-collapse" id="Plasmid_Construction"> | <div class="collapse multi-collapse" id="Plasmid_Construction"> | ||
</br></br> | </br></br> | ||
− | <p class="pcontent"> | + | <p class="pcontent"> |
− | <table class="card card-body"> | + | <ol> |
+ | <li>Digestion (vector)</li> | ||
+ | </ol> | ||
+ | </p> | ||
+ | <table class="card card-body PC1"> | ||
<tr> | <tr> | ||
<th>Plasmid</th> | <th>Plasmid</th> | ||
Line 195: | Line 200: | ||
</br> | </br> | ||
− | <p class="pcontent">2 | + | <p class="pcontent"> |
− | <table class="card card-body"> | + | <ol start="2"> |
+ | <li>Digestion (insert)</li> | ||
+ | </ol> | ||
+ | </p> | ||
+ | <table class="card card-body PC1"> | ||
<tr> | <tr> | ||
<th>Plasmid</th> | <th>Plasmid</th> | ||
Line 236: | Line 245: | ||
</br> | </br> | ||
− | <p class="pcontent">3 | + | <p class="pcontent"> |
− | + | <ol start="3"> | |
− | + | <li>Confirm the size of the digested product by gel electrophoresis.</li> | |
− | + | </br></br> | |
+ | <li>Gel purification of the target size.</li> | ||
+ | </br></br> | ||
+ | <li>Ligation</li> | ||
+ | </br> | ||
+ | </ol> | ||
+ | </p> | ||
− | + | <table class="card card-body"> | |
− | + | <tr> | |
− | + | <td>Vector (2 kbp)</td> | |
− | + | <td rowspan="2">molar ratio = 1:3</br>(can up to 1:10 depending on the | |
− | + | DNA sizes)</td> | |
− | + | </tr> | |
− | + | <tr> | |
− | + | <td>Insert (1.5 kbp)</td> | |
− | + | </tr> | |
− | + | <tr> | |
− | + | <td>Quick Ligase Reaction Buffer (2X)*</td> | |
− | + | <td>10 μl</td> | |
− | + | </tr> | |
− | + | <tr> | |
− | + | <td>Quick Ligase</td> | |
− | + | <td>1 μl</td> | |
− | + | </tr> | |
− | + | <tr> | |
− | + | <td>ddH2O</td> | |
− | + | <td>Up to 20 μl</td> | |
− | + | </tr> | |
− | + | </table> | |
− | + | </br> | |
− | + | <p class="pcontent"> | |
+ | <ol start="6"> | ||
+ | <li>Transform the product by heat shock.</li> | ||
+ | </br> | ||
+ | </ol> | ||
+ | </p> | ||
Revision as of 10:26, 29 September 2018