Template:SBB-Protocols Enz6

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4uL Miniprepped plasmid
4uL Miniprepped plasmid
1uL 10x NEB Buffer 2
1uL 10x NEB Buffer 2
-
0.5uL EcoRI
+
1uL EcoRI
-
0.5uL BamHI (for parts >250bp) or XhoI (for parts <250bp)
+
1uL BamHI (for parts >250bp) or XhoI (for parts <250bp)
FOR Righty Methylated Parts use Eco/Xho!
FOR Righty Methylated Parts use Eco/Xho!
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Revision as of 19:52, 23 June 2010

Set up the following 10uL reaction in a PCR tube:

4uL ddH2O
4uL Miniprepped plasmid
1uL 10x NEB Buffer 2
1uL EcoRI
1uL BamHI (for parts >250bp) or XhoI (for parts <250bp)
FOR Righty Methylated Parts use Eco/Xho!
  • Incubate at 37 on the thermocycler for 30 minutes
  • Run an analytical gel
  • Take a picture of the gel
  • Calculate the expected fragment sizes
  • Are the calculated sizes consistent with the bands on the gel?
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