Team:UNIPV-Pavia/Methods Materials/Digestion

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(New page: <h1>BioBrick digestion with restriction enzymes</h1> ''(estimated time: 3 hours)'' <br> <br> '''Materials needed:''' *'''Roche restriction enzymes thawed on ice''' *'''Roche buffer H''' *...)
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<h1>BioBrick digestion with restriction enzymes</h1>
<h1>BioBrick digestion with restriction enzymes</h1>
''(estimated time: 3 hours)''
''(estimated time: 3 hours)''

Revision as of 17:25, 22 June 2009

EthanolPVanimation.gif



Protocols


BioBrick digestion with restriction enzymes

(estimated time: 3 hours)

Materials needed:

  • Roche restriction enzymes thawed on ice
  • Roche buffer H
  • Pre-warmed at 37°C bath
  • Cut and gel-extracted vector
  • ddH2O


  • To open vectors:
    • a volume containing 1 µg of purified plasmid
    • 2 µl of buffer H
    • 1 µl of first enzyme
    • 1 µl of second enzyme
    • 20 µl final volume
    • incubate at 37°C for 3 hours
  • To excide fragments:
    • 20 µl of purified plasmid when <7 µg have been extracted. A volume containing 7 µg otherwise.
    • 2.5 µl of buffer H
    • 1 µl of first enzyme
    • 1 µl of second enzyme
    • 25 µl final volume
    • incubate at 37°C for 2 hours and 30 minutes


Retrieved from "http://2009.igem.org/Team:UNIPV-Pavia/Methods_Materials/Digestion"