AP-PCR QUESTION

Taek H. You tyou at postbox.acs.ohio-state.edu
Thu Jul 3 03:47:23 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?

Isn't there any different machine involved?
When I ran the same reactions in different machines, results were remarkably 
different.
In addition, many published stuffs were not reproducible until some twist in 
PCR.
 I am talking about a regular PCR, not AP-PCR. But, I think that might be 
the cause.



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