PCR and Genomic DNA contamination

teresa_mogul at my-deja.com teresa_mogul at my-deja.com
Mon Jun 14 21:07:39 EST 1999


Hi Dan

I've been running 16S universal PCR for bacteria. I have had a problem
getting my negative controls to be negative also. I have tried different
polymerases, dNTPs, waters etc. and each time I get at least 1 out of 8
of my negative controls positive. The only way that I have found to
combat this is to UV your mmix before PCR. I UV all ingredients but
not the polymerase, then add the polymerase and then dispense and do my
PCR. The way my negative controls are always negative. If you UV the
mmix with the polymerase in it it still works but I have found a
decrease in sensitivity. As I am trying to optimise my sensitivity this
is important to me.

Hope this helps

Teresa


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