Medium length PCR(2-5kb)?
hubahopp at gmx.de
Mon Jun 23 03:11:24 EST 2003
Basically it depends on what you want to do with your fragment. If you just
analyze it on a gel and do a restrictition digest, Taq should be
sufficient. If you want to clone it, use a prrofreading pol, adjust
extension time to 1 min per kb and still sequence your clones before
proceeding any further.
At 21:33 20.06.2003 +0100, Linan Chen wrote:
>hi, what enzyme do you recommend for medium length PCR(2-5kb) with good
>cost/effect? <2kb PCR can be reliably done by Taq(we use Taq from Promega),
>for >5kb LD-PCR kit from Roche or Takara is good. Because I want to use the
>enzyme to do mice genotyping(knockout mice), so it must be affordable and
>reliable. Or somebody has the homemade enzyme mix(Taq+pfu...). Thanks a lot.
>Chen, Linan Ph.D.
>Department of Neurobiology, Pharmacology and Physiology
>J.F. Knapp Research Center, Room R214
>924 E. 57th Street
>Chicago, Illinois 60637
>Email: lichen at drugs.bsd.uchicago.edu
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