Template:SBB-Protocols Enz2

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(New page: ===EcoRI/BamHI Digest of PCR Products===)
Current revision (15:51, 8 February 2011) (view source)
 
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===EcoRI/BamHI Digest of PCR Products===
===EcoRI/BamHI Digest of PCR Products===
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'''For PCR products, you will only digest a portion of your purified PCR product.'''
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*Set up the following reaction:
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  8uL of eluted PCR product
 +
  1uL of NEB Buffer 2
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  0.5uL EcoRI
 +
  0.5uL BamHI
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*Incubate at 37 degrees on the thermocycler for 1hr
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*Run an agarose gel, and melt with 600uL ADB buffer at 55 degrees.  ****NOTE:  If you are running short of time, this is an acceptable stopping point
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*If the DNA is shorter than 300bp, add 250uL of isopropanol and mix prior to loading it on the column

Current revision

EcoRI/BamHI Digest of PCR Products

For PCR products, you will only digest a portion of your purified PCR product.

  • Set up the following reaction:
 8uL of eluted PCR product
 1uL of NEB Buffer 2
 0.5uL EcoRI
 0.5uL BamHI
  • Incubate at 37 degrees on the thermocycler for 1hr
  • Run an agarose gel, and melt with 600uL ADB buffer at 55 degrees. ****NOTE: If you are running short of time, this is an acceptable stopping point
  • If the DNA is shorter than 300bp, add 250uL of isopropanol and mix prior to loading it on the column
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