PCR inhibitors - What are common pcr inhibitors (Mar/28/2006 )

I read somewhere that ethanol might inhibit PCR. My question is, if used to precipite DNA for extraction or when preserving the original sample. I had fish larvae samples that were preserved with DMSO the placed in ethanol 95%. This harms the DNA.

Do not damage the DNA but does not inhibit PCR either?

Thanks

if your samples are stored in EtOH before processing, this will not inhibit your PCR

what will inhibit your PCR is if there is residual ethanol in the final DNA sample that you will use as template; do you see the difference? if EtOH is present in the reaction, it will inhibit amplification. Your DNA can have all sorts of contact with EtOH prior to this step, as long as it is removed before the DNA is used as a template.

I was at a qPCR conference last week, and a couple of times they mentioned "testing for inhibitors". How is that accomplished? I was just trying to do a careful job purifying my RNA before I proceeded with anything, but I haven't come across any methods that described testing RNA or cDNA for the presence of inhibitors. Do you just do this using qPCR? Thanks!!

more than 2%DMSO may give non specific products. But for the rest, i agree with aimikins

back on my reply : over the night, i made a PCR with 5%DMSO with no pb. Actually, it was better than 0 to 2%...

Finnzyme's product sheet that comes with Phusion polymerase recommends testing a gradient of DMSO from 2-10%, for a really tricky PCR...but says more than 10% will inhibit.

I do not know if Taq can handle that much DMSO?

my boss do a 8% DMSO reaction fine.
He tests overnight a 12%DMSO Taq reaction.
I'll tell f he succeeded.