Team:Arizona State/Lab/Protocols/Restriction

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Protocols: Restriction


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Linearized Plasmid Backbone Restriction


  • Preparation of Master mix:
  • 5 uL NEB Buffer 4
  • 0.5 uL BSA
  • 0.5 uL EcoRI
  • 0.5 uL PstI
  • 0.5 uL DpnI
  • 18 uL dH2O
  • 4 uL linearized plasmid backbone + 4uL master mix in tube.
  • Digest at 37C for 30 minutes. Heat shock at 80C for 20 minutes.

[http://openwetware.org/wiki/Endy:Restriction_Digest Open WetWare Restriction]


  • Master mix (good for 8 restrictions)
  • 40 uL Buffer 2
  • 0.5 uL BSA
  • 296 uL ddH2O
  • 42.5 uL master mix + 300ng DNA + 1 uL of each restriction enzyme in tube.
  • Digest at 37C for 1 hour (or longer). Heat shock at 80C for 20 minutes.