Template:SBB-Protocols Enz6: Difference between revisions
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6.5 uL ddH2O | 6.5 uL ddH2O | ||
4uL Miniprepped plasmid | |||
1uL 10x NEB Buffer 2 | 1uL 10x NEB Buffer 2 | ||
0.5uL EcoRI | 0.5uL EcoRI |
Revision as of 15:21, 10 February 2010
Set up the following 10uL reaction in a PCR tube:
6.5 uL ddH2O 4uL Miniprepped plasmid 1uL 10x NEB Buffer 2 0.5uL EcoRI 0.5uL BamHI (for parts >250bp) or XhoI (for parts <250bp)
- 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?