Team:Groningen/protocols/Digestion

From 2013.igem.org

(Difference between revisions)
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   <tr>
   <tr>
     <td>MQ water</td>
     <td>MQ water</td>
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     <td ALIGN=CENTER>up to 20 &micro;l</td>
+
     <td>up to 20 &micro;l</td>
     <td></td>
     <td></td>
   </tr>
   </tr>
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   <tr>
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     <td>10x fast digestion buffer</td>
     <td>10x fast digestion buffer</td>
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     <td ALIGN=CENTER>2 &micro;l</td>
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     <td>2 &micro;l</td>
     <td>1x</td>
     <td>1x</td>
   </tr>
   </tr>
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   <tr>
     <td>restriction enzyme(s)</td>
     <td>restriction enzyme(s)</td>
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     <td ALIGN=CENTER>1 &micro;l</td>
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     <td>1 &micro;l</td>
     <td>1U</td>
     <td>1U</td>
   </tr>
   </tr>
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   <tr>
     <td>DNA</td>
     <td>DNA</td>
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     <td ALIGN=CENTER>4 &micro;l</td>
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     <td>4 &micro;l</td>
     <td>up to 1 &micro;g</td>
     <td>up to 1 &micro;g</td>
   </tr>
   </tr>
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   <tr>
   <tr>
     <td>MQ water</td>
     <td>MQ water</td>
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     <td ALIGN=CENTER>up to 20 &micro;l</td>
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     <td>up to 20 &micro;l</td>
     <td></td>
     <td></td>
   </tr>
   </tr>
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   <tr>
   <tr>
     <td>10x restriction digestion buffer</td>
     <td>10x restriction digestion buffer</td>
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     <td ALIGN=CENTER>2 &micro;l</td>
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     <td>2 &micro;l</td>
     <td>1x</td>
     <td>1x</td>
   </tr>
   </tr>
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     <td>restriction enzyme(s)</td>
     <td>restriction enzyme(s)</td>
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     <td ALIGN=CENTER>0.5 &micro;l</td>
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     <td>0.5 &micro;l</td>
     <td>1U</td>
     <td>1U</td>
   </tr>
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   <tr>
     <td>DNA</td>
     <td>DNA</td>
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     <td ALIGN=CENTER>2 &micro;l</td>
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     <td>2 &micro;l</td>
     <td>0.2-1.5 &micro;g</td>
     <td>0.2-1.5 &micro;g</td>
   </tr>
   </tr>

Revision as of 23:28, 4 October 2013

Restriction digestion

Fast digest

Materials:
  • MQ water
  • 1.5 ml tubes
  • Fast digestion buffer
  • Restriction enzymes
  • DNA
Reaction:
Compound 20 µl Final concentration
MQ water up to 20 µl
10x fast digestion buffer 2 µl 1x
restriction enzyme(s) 1 µl 1U
DNA 4 µl up to 1 µg

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

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


Normal digest

Materials:
  • MQ water
  • 1.5 ml tubes
  • Enzyme digestion buffer
  • Restriction enzymes
  • DNA
Reaction:
Compound 20 µl Final concentration
MQ water up to 20 µl
10x restriction digestion buffer 2 µl 1x
restriction enzyme(s) 0.5 µl 1U
DNA 2 µl 0.2-1.5 µg

When the reaction mix is ready, the content is mixed and incubated at 37°C for 2h.