Hello,

You can use Platinum Pfx DNA Polymerase since it is ideal for amplification of DNA fragments for high-fidelity PCR. High fidelity is provided by a proprietary enzyme preparation containing recombinant DNA polymerase from Thermococcus species KOD with proofreading activity.

I myself have used Hi-Fi-DNA polymerase and KOD Neo polymerase enzymes, luckily both maintained high fidelity.

Taq has a reputation for replication errors but suitable for routine amplification applications like genotyping or checking for the presence of an insert.

Best wishes!

Ok Thank you so much

I am trying to contact with toyobo company for KOD-Plus enzyme

Hi,

How many cycles are you doing? What is the size of the amplicon?

Thanks

Thanks a lot sir

I used 35 cycles and cloned 8 fragments of 1800, 1600,1400,1200,1000,800,600,400 upto 275bp

Hi,

You could try less cycles. Maybe you could try 30 or 32.This combine with another PFU enzyme should help.

Thanks so much

Angel

Hi,

It is important to use a Hi fidelity polymerase.

Thanks