Template:SBB-Protocols Enz1: Difference between revisions
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50uL eluted DNA | 50uL eluted DNA | ||
5.7uL NEB Buffer 2 | 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. | *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 | *Incubate the reaction at 37 degrees on the thermocycler | ||
*Proceed to another Zymo small fragment cleanup | *Proceed to another Zymo small fragment cleanup | ||
Latest revision as of 16:57, 23 June 2010
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
