Team:Newcastle/Restriction digests

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Restriction digestion

To cut DNA at specific base sequences using restriction enzymes

Procedures

  1. Make up 30 µl reaction mix as according:
  • 1 µl of restriction enzymes in total
  • 3 µl of 10x buffer
  • remainder is DNA/water
  1. Incubate at 37°C for 3 hours

Notes

  • No more than 10% of enzyme - solution contains glycerol which inhibits reaction
  • 10x buffer must be diluted to 1x i.e. 10% final volume
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