2 replies to this topic

[Guf]

Hello,

I need to amplify 7 kb sequence with high fidelity. We have three high-fidelity polymerases in our lab:


PfuUltra High-Fidelity DNA polymerase (from QuikChange II site directed mutagenesis kit, Agilent)

KAPA Hifi HotStart DNA polymerase (Kapabiosystems)

Q5 Hot Start High-Fidelity DNA polymerase (New England Biolabs)


Naturally, each manufacturer claims that his product is the best one. I have no experiences with HF polymerases so would it be possible to advise me which polymerase is really the best, please? Thank you.

[phage434]

My go-to polymerase is Q5. PfuUltra is somewhat tricky, no experience with Kappa.

[bigudukaz]

Kapa is no good compared to the other ones. best options - Q5 (NE biolabs) and Phusion high fidelity DNA polimerase (from ThermoFisher) they are both very good, but it depends on the system of the reaction which one is better. but for 7kb i guess Phusion is better.