PCR from genomic DNA

krishnan at BORCIM.WUSTL.EDU krishnan at BORCIM.WUSTL.EDU
Wed Apr 6 14:54:46 EST 1994


Hi!
Could somebody tell me how to PCR-amplify a 3 kb piece of DNA from total human
genomic DNA or from total yeast DNA (from a strain containing the respective
YAC clone)?  I tried 1 microgram template, 50 pmol primers, 200 micromole dNTPS
2.5 mM Mg and 1 unit Amplitaq.  The cycling profile was: 96 C 2 min 1 cycle,
94 C 1 min, 55 C 1 min, 72 C 3 min for 30 cycles. I didn't get any product as
determined by EtBr staining. Thanx in advance for any pointers.
Raja
krishnan at borcim.wustl.edu



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