UDG in qPCR - (Aug/09/2012 )
I would like if anyone can make any comments about using UDG in qPCR. Improves qPCR results? Any experience with it?
Thank in advance
UDG degrades templates containing dUTP nucleotides incorporated in DNA. At the start of the reaction UDG degrades any contamination from previous reactions that contained dUTP in reaction mix, leaving only the native template.
UDG is used as standard in routine and diagnostic qPCR applications, or when you have repeated problems with contamination by previous PCR producs.
UDG won't improve quality of qPCR results anyhow apart from this specific contamination, nor it will prevent cross-contamination with other samples, that are not PCR products amplified with dUTP.
Most commercial master mixes already contain dUTP by default with possibility to buy UDG for routine treatment or in case of contamination.
Never need to use that, I think most problems with product contamination can be avoided by strictly separating the places, materials and tools where you set up and analyse results (though in case of real-time PCR there is mostly no need to ever open used tube or plate, this was more of a case in classic PCR).
I think in ordinary lab if you keep this rule, you don't need to spend money on UDG treatments.