What do you get when you cross a Pyrococcus DNA polymerase with a dsDNA binding domain? …a highly processive, high fidelity, lighting fast PCR work-horseThose Finnish wizards at Finnzymes have made that work-horse a reality with their Phusion polymerase. I had been meaning to try out this enzyme for quite a while, but never got around to it, then a problematic 11 kb amplification brought it back to mind. Phusion is a next-generation polymerase whose incorporated dsDNA binding domain increases it’s processivity by 10-fold, and it’s extension rate to a mind-boggling 1kb/15 seconds.. pfu is left in the dust!
But enough of the hype – does it work? Well after spending a frustrating 2 weeks messing around with Pfu, Taq, temperature gradients I got a beautiful amplified band at my first attempt with Phusion. Not only that, but the reduction in the extension time from 22 minutes to 5.5 minutes (I went for the upper end of the recommended extension time) certainly helped to get the reaction done faster, and me to get home earlier.
If you have tried out the Phusion polymerase, please feel free to post your comments below. If you liked this article please consider subscribing to my article feed. Please note that this site has no association whatsoever with Finnzymes or NEB (or any other biotech company for that matter!)
Antibiotics are used in a wide range of techniques in molecular biology. My aim with this post is to provide an easy reference to some of the main ones used in molecular biology, their mechanisms, range and working concentrations. I hope you will find it useful. Your Personal Antibiotics Reference Guide *Abbreviations: Gm(+/-)=Gram positive/negative; My=mycoplasma; […]
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