Difference between revisions of "Template:SBB-Protocols Enz3"

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(BglII Digest of EIPCR Products)
 
Line 4: Line 4:
 
   8.5uL of eluted PCR product
 
   8.5uL of eluted PCR product
 
   1uL of NEB Buffer 2
 
   1uL of NEB Buffer 2
   0.5uL BglII
+
   1uL BglII
 
*Incubate at 37 degrees on the thermocycler for 1hr
 
*Incubate at 37 degrees on the thermocycler for 1hr
 
*Run an agarose gel, cut out the band, and melt with 600uL ADB buffer at 55 degrees
 
*Run an agarose gel, cut out the band, and melt with 600uL ADB buffer at 55 degrees
 
*Proceed with remainder of Zymo gel purification protocol
 
*Proceed with remainder of Zymo gel purification protocol

Latest revision as of 15:54, 23 June 2010

BglII Digest of EIPCR Products

For EIPCR products, you will only digest a portion of your purified PCR product. This procedure is almost identical to the digestion of a PCR product with EcoRI/BamHI, but you'll use BglII instead of EcoRI/BamHI.

  • Set up the following reaction:
 8.5uL of eluted PCR product
 1uL of NEB Buffer 2
 1uL BglII
  • Incubate at 37 degrees on the thermocycler for 1hr
  • Run an agarose gel, cut out the band, and melt with 600uL ADB buffer at 55 degrees
  • Proceed with remainder of Zymo gel purification protocol