Amplifying converted DNA is one of the trickiest steps in any Bisulfite conversion based protocol. A mistake here could stop your study cold if the amplification fails, or worse yet, you might be introducing errors that you wonât see until youâve spent hours combing through useless sequencing data. Pfu-Turbo Cx Hotstart DNA Polymerase from Agilentâs Stratagene Division, takes care of those issues by combining proofreading activity for low error rates, with the ability to efficiently read through Uracil without stalling for better amplification of converted DNA templates.
This Pfu Allows U-Turns
Researchers normally have used Taq polymerases with bisulfite converted DNA because they can read through Uâs (uracil), while proofreading enzymes, like regular Pfu enzymes stall, leaving you with little product. Ah, but Pfu-Turbo Cx Hotstart isnât your normal PfuâŚItâs a mutated version thatâs been developed to fly through Uâs without a second thought, so you wonât have to sacrifice Pfuâs proofreading for Taqâs yield.
Proofreading That Makes Spell-Check Jealous
With that pesky uracil issue settled, the best reason for using a Pfu polymerase is the low error rates. Sure Taqpolymerases can run off a ton of amplification products, but unlike Pfu (with an error rate of just 1 in 780,000) they can be a little sloppy, creating errors about once every 125,000 bases. (Nucleic Acids Research, September 1996) Pfu-Turbo Cx Hotstartâs proofreading feature will have your sequencer calling more letters right than a 12yr-old spelling bee champ, which really pays off when you are thinking of taking on a whole-genome study.
Let your Sequencing Data Shine
Pfu Turbo Cx Hotstart DNA Polymerase is tailor made for todayâs powerful combinations of bisulfite and next generation sequencing (NGS) methods like Reduced Representation Bisulfite Sequencing (RRBS). Reducing amplification errors early keeps them from spiraling out of control later on when youâre spending lots of time and money on sequencing runs.
Make sure that Pfu-Turbo Cx Hotstart is a part of your next bisulfite sequencing run. Check out the Agilent websiteto get your hands on some.