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?