stylized illustration representing PCR

Phire Hot Start II DNA Polymerase

A fast and robust enzyme is born by fusing a DNA polymerase to a small dsDNA-binding protein
  • Frustrated by failed PCR reactions?
  • Wondering what reaction inhibitors caused your luck-luster results?

Inhibition of a PCR reaction is often caused by substances that are purified along with the template DNA (or RNA in the case of RT-PCR reaction). The inhibiting substances interact with the DNA molecule and prevent the polymerase from binding. Standard DNA polymerases dissociate frequently and easily from the DNA template, allowing inhibitors to come in and interact with the DNA molecule. Polymerases are then hindered from binding to the template again, resulting in poor or no amplification. Pondering this, our Thermo Fisher Scientific R&D scientists asked themselves “What happens if you improve the affinity of the DNA polymerase to the DNA, by adding a double stranded (ds) DNA-binding domain to it?"

And, that was how the Thermo Scientific Phire Hot Start II DNA Polymerase was born. This polymerase was created by fusing a polymerase enzyme and a small dsDNA- binding protein. The dsDNA- binding domain keeps the polymerase on the DNA template and decreases its chances of falling off. This in turns reduces the ability of inhibitors to hinder its binding to DNA. In addition, since Phire polymerase stays on the DNA, polymerization occurs faster because no time is wasted waiting for the DNA polymerase to latch back on to the template. It's actually possible to skip the DNA purification step altogether and perform a direct PCR, since contaminating inhibitors from the sample can't stop our Phire DNA polymerase!

To summarize

Phire Hot Start II DNA Polymerase is a very robust DNA polymerase, which can work in tough conditions, going faster (10-15 s/kb!) and further (up to 20 kb!) and giving higher yields – making your routine PCR a piece of cake!