Template:SBB-Protocols Enz1
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EcoRI/BamHI Digest of Wobble Products
For wobble products, you will digest the entire extension reaction-worth of DNA Since you started with 50uL of extension reaction, you should now have 50uL of eluted DNA in water.
- Set up the following reaction in a PCR tube:
50uL eluted DNA 5.7uL NEB Buffer 2 0.5uL EcoRI 0.5uL BamHI
- Mix thoroughly by slamming the tube upside down on the table, and then a quick spin to move the liquid to the bottom of the tube.
- Incubate the reaction at 37 degrees on the thermocycler
- Proceed to another Zymo small fragment cleanup