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Team:Tuebingen/Notebook/Protocols/pcr
From 2013.igem.org
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PCR | PCR | ||
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| + | <h3>Reagents</h3> | ||
| + | <table border="0"> | ||
| + | <colgroup> | ||
| + | <col width="80"> | ||
| + | <col width="300"> | ||
| + | </colgroup> | ||
| + | |||
| + | <tr> | ||
| + | <td style="text-align: center">39.5 µL</td> | ||
| + | <td>Aqua dest.</td> | ||
| + | </tr> | ||
| + | |||
| + | <tr> | ||
| + | <td style="text-align: center">2.5 µL</td> | ||
| + | <td>dNTPs (200 µM each)</td> | ||
| + | </tr> | ||
| + | |||
| + | <tr> | ||
| + | <td style="text-align: center">1 µL</td> | ||
| + | <td>DNA (c = 100 ng/µL)</td> | ||
| + | </tr> | ||
| + | |||
| + | <tr> | ||
| + | <td style="text-align: center">5.0 µL</td> | ||
| + | <td>Taq-Buffer</td> | ||
| + | </tr> | ||
| + | |||
| + | <tr> | ||
| + | <td style="text-align: center">0.5 µL</td> | ||
| + | <td>Forward primer</td> | ||
| + | </tr> | ||
| + | |||
| + | <tr> | ||
| + | <td style="text-align: center">0.5 µL</td> | ||
| + | <td>Reverse primer</td> | ||
| + | </tr> | ||
| + | |||
| + | <tr> | ||
| + | <td style="text-align: center">1 µL</td> | ||
| + | <td>Taq/Pfu polymerases (9+1)</td> | ||
| + | </tr> | ||
| + | </table> | ||
| + | |||
| + | <p> </p> | ||
| + | |||
| + | <h3>Procedure</h3> | ||
| + | <p>Mix all reagents in the given order (Important: add polymerases last!) and transfer in one 0.2 mL Eppendorf-tube ("PCR-tube"). Run cycler according to your requirements.</p> | ||
| + | |||
| + | <p><b>This protocol yields exactly one single reaction mixture!</b> Scale linearly for multiple reaction mixtures and keep mixture on ice or even better at -20 °C for best results.</p> | ||
| + | |||
| + | |||
| + | <a href="https://2013.igem.org/Team:Tuebingen/Notebook/Protocols" style="font-size: 18px">Back to Protocols</a> | ||
</div> | </div> | ||
Latest revision as of 12:09, 4 October 2013
PCR
Reagents
| 39.5 µL | Aqua dest. |
| 2.5 µL | dNTPs (200 µM each) |
| 1 µL | DNA (c = 100 ng/µL) |
| 5.0 µL | Taq-Buffer |
| 0.5 µL | Forward primer |
| 0.5 µL | Reverse primer |
| 1 µL | Taq/Pfu polymerases (9+1) |
Procedure
Mix all reagents in the given order (Important: add polymerases last!) and transfer in one 0.2 mL Eppendorf-tube ("PCR-tube"). Run cycler according to your requirements.
This protocol yields exactly one single reaction mixture! Scale linearly for multiple reaction mixtures and keep mixture on ice or even better at -20 °C for best results.
Back to Protocols
