I am using Deep Vent polymerase to amplify a region in a GOI in genomic DNA. I initally optimised my reaction conditions on plasmid DNA containing my GOI, but when I tried to amplify my GOI in genomic DNA I got no product.
The plasmid DNA and genomic DNA templates were diluted such that they have equal amounts of GOI target i.e:
1.07 pg pDNA = 160, 000 copies
and 1 ug gDNA = 160, 000 copies
The plasmid alone amplifies well, however when spiked into null genomic DNA I get no product! What is it about the background of genomic DNA that could be causing inhibition for Deep Vent??? I have tried this reaction with regular taq and it works perfectly. Could the exonuclease activity of Deep Vent have something to do with it?
Any suggestions or comments would be greatly appreciated!
Edited by jessh, 10 February 2010 - 08:21 PM.