Team:Tuebingen/Notebook/Protocols/3a-assembly

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<a href="https://2013.igem.org/Team:Tuebingen/Notebook/Protocols" style="font-size: 16px">Back to Protocols</a>
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<a href="https://2013.igem.org/Team:Tuebingen/Notebook/Protocols" style="font-size: 18px">Back to Protocols</a>
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<p>&nbsp;</p>
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</div>
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<h3>Reagents</h3>
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<table border="0">
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  <colgroup>
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    <col width="80">
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    <col width="300">
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  </colgroup>
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  <tr>
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    <td style="text-align: center">250 - 300 ng</td>
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    <td>Plasmid DNA</td>
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  </tr>
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  <tr>
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    <td style="text-align: center">2.5 µL</td>
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    <td><a href="https://www.neb.com/products/b7002-nebuffer-2">10x NEBuffer 2</a></td>
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  </tr>
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  <tr>
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    <td style="text-align: center">0.25 µL</td>
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    <td>100x BSA</td>
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  </tr>
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  <tr>
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    <td style="text-align: center">0.5 µL</td>
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    <td>Restriction enzyme A</td>
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  </tr>
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  <tr>
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    <td style="text-align: center">0.5 µL</td>
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    <td>Restriction enzyme B</td>
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  </tr>
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  <tr>
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    <td style="text-align: center">fill up to 25 µL</td>
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    <td>Aqua dest.</td>
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  </tr>
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</table>
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<p>&nbsp;</p>
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<h3>Procedure</h3>
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<p>Dissolve all reagents in 1000 mL Aqua dest., adjust pH to 7.0 (with NaOH). Sterilize by autoclaving. After autoclaving add 5 mL MgCL2 (c = 2 M).</p>
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Revision as of 00:57, 4 October 2013

Return to iGEM Main Page.

3A-Assembly
Back to Protocols

 

Reagents

250 - 300 ng Plasmid DNA
2.5 µL 10x NEBuffer 2
0.25 µL 100x BSA
0.5 µL Restriction enzyme A
0.5 µL Restriction enzyme B
fill up to 25 µL Aqua dest.

 

Procedure

Dissolve all reagents in 1000 mL Aqua dest., adjust pH to 7.0 (with NaOH). Sterilize by autoclaving. After autoclaving add 5 mL MgCL2 (c = 2 M).