Template:SBB-Protocols Enz1: Difference between revisions
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===EcoRI/BamHI Digest of Wobble Products=== | ===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 |
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