Template:SBB-Protocols Enz1

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(New page: ===EcoRI/BamHI Digest of Wobble Products===)
Current revision (19:57, 23 June 2010) (view source)
 
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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
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  5.7uL NEB Buffer 2
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  0.5uL EcoRI
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  0.5uL BamHI
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*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.
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*Incubate the reaction at 37 degrees on the thermocycler
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*Proceed to another Zymo small fragment cleanup

Current revision

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
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