Template:SBB-Protocols Enz3: Difference between revisions
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===BglII Digest of EIPCR Products=== | ===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 | |||
0.5uL 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 | |||
Revision as of 21:58, 3 February 2009
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 0.5uL 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
