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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