PCR directly from phage

Ron Kagan rkagan at ewald.mbi.ucla.edu
Tue Sep 13 19:30:50 EST 1994


In article <hxm8.69.000F93AA at po.cwru.edu> Harry Menegay, hxm8 at po.cwru.edu
writes:
>I'm trying to PCR out of a phage cDNA library to determine
>if my gene of interest is located in the library before I
>go ahead and screen the library.  My question is how many 
>phage can you put in a PCR reaction before the reaction dies
>due to too much protein from all the phage?  Anyone tried this?
>
>Thanks for any help
>Harry

I successfully cloned a gene from a cDNA library in Lambda Zap, using 2
ul of the library (1X10^9 pfu/ml) per 50 ul PCR reaction.  Just heat the
reaction mix at 98C for 3 min. before adding the Taq and then cycle as
usual.


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"My Kindness is not random, and nothing senseless was ever beautiful."

                                                 - J. Zabriskie
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Ron Kagan
rkagan at ewald.mbi.ucla.edu



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