PCR troubleshooting

Terese M Barta-1 barta001 at STAFF.TC.UMN.EDU
Wed Jun 30 23:05:10 EST 1993

Does anyone out there know if RNA in a DNA sample can interfere with
PCR amplication? Lately, I have had a total lack of success getting product
(500 bp) from chromosomal DNA although I had been successful previously.
My reagents and procedure are all the same as before and the positive
control (plasmid template) works fine.The only apparent difference
is that I am using a different batch of bacterial genomic DNA.  The "old"
batch was CsCl-purified but my latest batch is minipreped (and not RNased).

Someone in my lab mentioned that RNA possibly interferes with PCR with
chromosomal DNA.If you have any experiences or refs that support this, I'd appreciate
hearing from you. Please reply directly. I will post a summary of
responses if anything turns up. Thanks in advance.

T. Barta
Univ. of MN

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