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