Suggestions - PCR from Phage Isolates
Tori D. Williams
twilliam at biomed.med.yale.edu
Thu Feb 15 11:11:53 EST 1996
I have taken a lambda phage library through 2 rounds of screening. I
have over 80 clones that I would like to further screen by PCR. I have
developed primers that appear to give me product using total human DNA -
although not as clean as I would like - but I can't get anything from a
mix of the phage eluates. My eluates are in SM and my latest PCR
conditions have been 94, 52 and 72 C for 1 min each x 35. I have tried
adding and not adding extra Mg++. I am getting lots of primer dimers
though. I would appreciate any suggestions. I have found one protocol
that involves amplifying the phage in microtiter plates. I will be
trying that tomorrow.
Thanks,
Tori
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