Fast and Robust Routine PCR

Inhibition of a PCR reaction is often caused by substances that are present in the samples or are carried over from the purification process. These inhibitors often interact with the reaction components and the DNA-enzyme complexes formed during PCR. Standard PCR enzymes like Taq DNA polymerase dissociate frequently and easily from the DNA template, allowing inhibitors to come in and interact with the DNA molecules and the enzymes. The enzymes are then hindered from binding to the templates again, resulting in poor or no amplification. Pondering this, Thermo Fisher Scientific scientists asked themselves “How about if you improve the affinity of the DNA polymerase to the DNA, by adding a double-stranded (ds) DNA-binding domain to the polymerase?"

And, that was how the Thermo Scientific Phire Hot Start II DNA Polymerase was born. This polymerase was created by fusing a polymerase and a small dsDNA-binding protein. The dsDNA- binding domain keeps the polymerase on the DNA template longer and decreases its chances of falling off. This phenomenon allows faster polymerization while enabling higher tolerance to the inhibitors. Thanks to this innovation, you can skip the DNA purification step altogether and perform a direct PCR, since inhibitors in the sample can't match our Phire DNA polymerase.

In summary, Phire Hot Start II DNA Polymerase is a very robust DNA polymerase. Compared to Taq DNA polymerase, it can work through tough reaction conditions; it can go faster (10–15 sec/kb) and further (7.5 kb); and it can give higher yields of target amplicons. Overall, it can make your hot-start PCR easier and more efficient!