Team:Groningen/protocols/Digestion

From 2013.igem.org

(Difference between revisions)
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     <td>DNA</td>
     <td>DNA</td>
     <td>4&micro;l</td>
     <td>4&micro;l</td>
-
     <td>up to 1&micro;l</td>
+
     <td>up to 1&micro;g</td>
   </tr>
   </tr>
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<br>
<br>
<br>Reference: http://www.thermoscientificbio.com/uploadedFiles/Resources/fast-digestion-dna.pdf
<br>Reference: http://www.thermoscientificbio.com/uploadedFiles/Resources/fast-digestion-dna.pdf
 +
 +
 +
<h2>Normal digest</h2>
 +
 +
<h3>Materials:</h3>
 +
<ul type="square">
 +
<li>MQ water</li>
 +
<li>1.5ml tubes</li>
 +
<li>Enzyme digestion buffer</li>
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<li>Restriction enzymes</li>
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<li>DNA</li>
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</ul>
 +
 +
<h3>Reaction:</h3>
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<table border="1">
 +
  <tr>
 +
    <th>Compound</th>
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    <th>20&micro;l</th>
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    <th>Final concentration</th>
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  </tr>
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 +
  <tr>
 +
    <td>MQ water</td>
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    <td>up to 20&micro;l</td>
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    <td></td>
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  </tr>
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 +
<tr>
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    <td>BSA</td>
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    <td>0.5&micro;l</td>
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    <td></td>
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  </tr>
 +
 +
  <tr>
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    <td>10x restriction digestion buffer</td>
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    <td>2.5&micro;l</td>
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    <td>1x</td>
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  </tr>
 +
 +
  <tr>
 +
    <td>restriction enzyme(s)</td>
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    <td>0.5&micro;l</td>
 +
    <td>1U</td>
 +
  </tr>
 +
 +
  <tr>
 +
    <td>DNA</td>
 +
    <td>2&micro;l</td>
 +
    <td>up to 200ng</td>
 +
  </tr>
 +
 +
</table>
</div>
</div>

Revision as of 14:34, 27 July 2013

Digestion

Fast digest

Materials:

  • MQ water
  • 1.5ml tubes
  • Fast digestion buffer
  • Restriction enzymes
  • DNA

Reaction:

Compound 20µl Final concentration
MQ water up to 20µl
10x fast digestion buffer 2µl 1x
restriction enzyme(s) 1µl 1U
DNA 4µl up to 1µg

When the reaction mix is ready, the content is mixed an incubated at 37°C for 1h.

Reference: http://www.thermoscientificbio.com/uploadedFiles/Resources/fast-digestion-dna.pdf

Normal digest

Materials:

  • MQ water
  • 1.5ml tubes
  • Enzyme digestion buffer
  • Restriction enzymes
  • DNA

Reaction:

Compound 20µl Final concentration
MQ water up to 20µl
BSA 0.5µl
10x restriction digestion buffer 2.5µl 1x
restriction enzyme(s) 0.5µl 1U
DNA 2µl up to 200ng