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HotStarTaq PCR Handbook


PCR stock solutions can also be decontaminated using UV light. This method is laborious, however, and its efficiency is difficult to control and cannot be guaranteed. We recommend storing solutions in small aliquots and using fresh aliquots for each PCR.

Contamination by PCR product carry-over can be eliminated by using the commercially available uracil-N-glycosylase (UNG). The procedure involves substituting dUTP for dTTP in the PCR setup and treating all PCR mixtures with UNG prior to PCR amplification. As a result, any PCR product containing dUTP carried over from previous rounds of amplification is destroyed by cleavage during the initial incubation step that activates HotStarTaq DNA Polymerase.

Another approach to preventing amplification of contaminating DNA is to treat individual reaction mixtures with DNase I or restriction enzymes that cut between the binding sites of the amplification primers used, before adding the template DNA sample.

* Most commercial bleach solutions are approximately 5.25% sodium hypochlorate. Sodium hypochlorate is an irritant and should be handled with caution.

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