Team:Tuebingen/Notebook/Protocols/colony-pcr

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Colony-PCR
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<h3>Reagents</h3>
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    <td style="text-align: center">0.5 µL</td>
    <td>MgCl2 (c = 25 mM)</td>
  </tr>

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    <td style="text-align: center">1.0 µL</td>
    <td>10x Taq Polymerase Buffer</td>
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    <td style="text-align: center">0.1 µL</td>
    <td>dNTPs (2.5 mM each)</td>
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    <td style="text-align: center">0.1 µL</td>
    <td>Oligo forward</td>
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    <td style="text-align: center">0.1 µL</td>
    <td>Oligo reverse</td>
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    <td style="text-align: center">7.7 µL</td>
    <td>Aqua dest.</td>
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    <td style="text-align: center">0.1 µL</td>
    <td>Taq Polymerase (5 U/µL)</td>
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<p>&nbsp;</p>

<h3>Procedure</h3>
<ol>
  <li>Put some PCR-tubes on ice.</li>
  <li>The given volumes are for one single reaction of 15 µL. When checking more than one colony, repare a master mix for all colonies. Keep that master mix on ice all the time!</li>
  <li>Transfer 10 µL of master mix in every ice-cold PCR-tube (1 PCR-tube per colony).</li>
  <li>Usa a pipet tip in order to pick up a colony from a fresh plate and transfer the colony into the reaction mixture inside a PCR-tube.</li>
  <li>Smear the tip over a new LB plate + antibiotic in order to create new colonies.</li>
  <li>Run PCR and control success via <a href="https://2013.igem.org/Team:Tuebingen/Notebook/Protocols/gelelectrophoresis">gelelctrophoresis</a>.</li>
  <li>Store plates of successful colony-PCRs at 4 °C.
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<p>&nbsp;</p>
<a href="https://2013.igem.org/Team:Tuebingen/Notebook/Protocols" style="font-size: 18px">Back to Protocols</a>
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