PCR of phage library

Steven Goldberg goldberg at bms.com
Mon Sep 11 06:50:08 EST 1995


I am trying to clone a gene from a porcine cDNA library in gt10 using 
PCR.  The insert is ca. 1700 bp and its DNA sequence is known.  So far I 
have been able to amplify fragments of 500 and 600 bp from both ends of 
the cDNA using an aliquot of the denatured phage library as template, 
which is exactly as predicted.  However, I can't get the full-length 
gene to amplify, although the primers representing its NH2 and COOH 
termini are the same used in the successful experiments described above.
What sort of modifications might be useful to obtain the longer PCR 
product I'm looking for?

Thanks for any ideas.

Steve 





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