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Phage as a template for PCR?

Namjin Chung chung006 at mc.duke.edu
Thu Dec 7 14:15:11 EST 1995


Dear Netters,

I have gotten no products from PCR.  To clone out a gene from C. elegans cDNA library, I 
got cDNA library  in lambda phage which had been stored for a year or two in a refrigerator 
from other lab.  I tried different sets of conditions for PCR: i) with/without  
heat-pretreatment of phage solution for 3 min at 95 or 100 degree; ii) a range of Mg++ 
concentration; iii) different anealing temperature.

I got no band at all from agarose gel electrophoresis analysis, and I have began to suspect 
the DNA template.  I don't know if DNA contained in phage would be stable for over a 
year, and if my heat pretreatment efficiently stripped off phage proteins to expose DNA.  If 
anyone has experience in this kind of PCR, please give some advice to me at 
chung006 at mc.duke.edu.  Thanks in advance.

Namjin Chung
Program in Molecular Cancer Biology
Department of Cell Biology 
Duke University Medical Center
e-mail: chung006 at mc.duke.edu





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