PCR sequencing problems
hbbio305 at email.csun.edu
Mon Oct 20 12:34:11 EST 1997
I have been sequencing short to medium lenght PCR products successfully
(on a 377 ABI) up until a few weeks ago. I use shrimp alkaline
phosphatase and exonuclease I as a cleanup.
Recently all PCR sequencing has come to a halt. I get only short blips
on the gel image. As well, the controls have much higher background,
and more N's than normal.
I have tried increasing the DNA and ethanol precipitation of the DNA to
remove the inactive enzymes, to no avail.
Has anyone else had this problem?
Are there any suggestions?
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