Template:SBB-Protocols Enz1

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(New page: ===EcoRI/BamHI Digest of Wobble Products===)
(EcoRI/BamHI Digest of Wobble Products)
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===EcoRI/BamHI Digest of Wobble Products===
===EcoRI/BamHI Digest of Wobble Products===
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'''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.
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*Set up the following reaction in a PCR tube:
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  50uL eluted DNA
 +
  5.7uL NEB Buffer 2
 +
  1uL EcoRI
 +
  1uL 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
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*Proceed to another Zymo small fragment cleanup

Revision as of 23:48, 3 February 2009

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
 1uL EcoRI
 1uL 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
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