Team:Newcastle/Restriction digests


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Restriction digestion

Materials required

  • Eppendorf tubes
  • Pipettes
  • Appropriate DNA/plasmid
  • Appropriate restriction enzymes
  • 10X buffer
  • Water


Add the following as mentioned below to make up to a final volume of 20 µl of reaction mix:

  1. 15 µl of DNA/plasmid
  2. 1 µl of restriction enzyme 1
  3. 1 µl of restriction enzyme 2
  4. 2 µl of 10X buffer
  5. 1 µl of water

Incubate the digestion mixture at 37°C for 3 hours


  • No more than 10% of enzyme should be used for a single reaction - glycerol inhibits reaction
  • 10x buffer must be diluted to 1x i.e. 10% final volume.

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