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

From 2013.igem.org

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<p>&nbsp;</p>
<p>&nbsp;</p>
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<h2>Digestion</h2>
<h3>Reagents</h3>
<h3>Reagents</h3>
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   <tr>
   <tr>
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     <td style="text-align: center">250 - 300 ng</td>
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     <td style="text-align: center">20.0 g</td>
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    <td>Plasmid DNA</td>
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     <td>Trypton</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>
   </tr>
   <tr>
   <tr>
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     <td style="text-align: center">0.5 µL</td>
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     <td style="text-align: center">5.0 g</td>
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     <td>Restriction enzyme A</td>
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     <td>Yeast extract</td>
   </tr>
   </tr>
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   <tr>
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     <td style="text-align: center">0.5 µL</td>
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     <td style="text-align: center">0.5 g</td>
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     <td>Restriction enzyme B</td>
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     <td>NaCl</td>
   </tr>
   </tr>
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     <td style="text-align: center">to 25 µL</td>
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     <td style="text-align: center">10 mL</td>
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     <td>Aqua dest.</td>
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     <td>KCl (c = 250 mM)</td>
   </tr>
   </tr>
</table>
</table>

Revision as of 01:35, 4 October 2013

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3A-Assembly
Back to Protocols

 

Digestion

Reagents

20.0 g Trypton
5.0 g Yeast extract
0.5 g NaCl
10 mL KCl (c = 250 mM)

 

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).


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