DD/PCR products hang on wells

[babalola at MAIL.SAS.UPENN.EDU]

Hi there netters:

I do the differential display RT/PCR routinely. However, there has been a
menacing problem that I now am determined to solve.

The problem is what seems to be a random failure of pcr even within
replicate RT product templates. I just did an assay last week and 7 out of
my 18 samples (6 triplicates = 18 samples) apparently failed.

However, I can not say for certain that the pc reaction actually failed or
that something else is going on here. This is because the gel lanes on
which I ran the failed reactions always had intense radioactive signals
hanging at the wells while the "good reactions" never had this amount at
the wells.

I have begun to test the origin of this problem and find solutions to them
but, I will like the input of people who use this assay or who have an idea
of what may be going on here as I go along the troubleshooting path.


Thank you.

G. Lanre Babalola (Babalola at mail.sas.upenn.edu)
University of Pennsylvania
Philadelphia