PCR PROBLEMS

anitha anitha at EARTHLINK.NET
Thu Mar 25 07:35:56 EST 1999


Hello,

        I am trying to characterize a dominat mutation and I have some
problems in PCR sequencing. When I use the forward primer for sequencing my
PCR product, I get a large number of mutations. This is seen in all the
affected patients. When I use the reverse primer, these mutations disappear.
Can anybody explain this?

        Is it possible that, while doing the PCR (I use the same primers for
sequencing also) the mutant chromosome is preferentialy amplified with the
forward primer???


Please let me know your thoughts.


Anitha

anitha at earthlink.net





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