Template:SBB-Protocols Enz6: Difference between revisions
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4uL Miniprepped plasmid | 4uL Miniprepped plasmid | ||
1uL 10x NEB Buffer 2 | 1uL 10x NEB Buffer 2 | ||
.5uL EcoRI | |||
.5uL BamHI (for parts >250bp) or XhoI (for parts <250bp) | |||
FOR Righty Methylated Parts use Eco/Xho! | FOR Righty Methylated Parts use Eco/Xho! | ||
</pre> | </pre> |
Revision as of 16:55, 23 June 2010
Set up the following 10uL reaction in a PCR tube:
4uL ddH2O 4uL Miniprepped plasmid 1uL 10x NEB Buffer 2 .5uL EcoRI .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?