From 2011.igem.org
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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.
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