AP-PCR QUESTION

Jenny Williams Jenova at microbes.demon.co.uk
Sun Jun 29 13:47:50 EST 1997


I have recently completed a project on molecular typing using Arbitrary-
Primer PCR (AP-PCR). This technique uses a single arbitrary primer in
each PCR assay. I used random primers based on published sequences which
had previously been applied successfully in the typing of Clostridium
difficile. I followed the published protocols carefully, but was unable
to get reproducible results.  I then combined the same two random
primers that I had previously used in single primer PCR assays, in the
same reaction tube. The assay conditions were identical to that of the
single primer assays, with the exception that 2 primers were now taking
part in the reaction. Also, the total amount of primer in the reaction
tube was doubled. This combination produced reproducible results. Also,
the resulting DNA profile produced after gel electrophoresis was
different from the sum of the DNA profiles of the single primer assays.
Can anyone explain these phenomena?

Regards 
-- 
Jenny Williams



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