Team:Groningen/protocols/Digestion

From 2013.igem.org

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<h1>Restriction digestion</h1>
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<h2>Restriction digestion</h2>
<h3>Fast digest</h3>
<h3>Fast digest</h3>

Revision as of 11:24, 28 July 2013

Restriction 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 and 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
10x restriction digestion buffer 2µl 1x
restriction enzyme(s) 0.5µl 1U
DNA 2µl 0.2-1.5µg

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