Template:SBB-Protocols Enz6: Difference between revisions
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0.5uL EcoRI | 0.5uL EcoRI | ||
0.5uL BamHI (for parts >250bp) or XhoI (for parts <250bp) | 0.5uL BamHI (for parts >250bp) or XhoI (for parts <250bp) | ||
FOR Righty Methylated Parts use Eco/Xho! | |||
</pre> | </pre> | ||
Revision as of 17:56, 18 June 2010
Set up the following 10uL reaction in a PCR tube:
4uL ddH2O 4uL Miniprepped plasmid 1uL 10x NEB Buffer 2 0.5uL EcoRI 0.5uL 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?