Hi,

Short answer is I haven't done 20 kb PCR, but I have heard Takara Bio LA Taq recommended for 23 kb PCR from genomic DNA, so could be worth a go.

Chris

I´ve used Takara Bio LA Taq for mtDNA (16 Kb) and it works better than others.

Some of the suggestions from this discussion might be useful for you:

https://www.researchgate.net/post/Does_anybody_know_a_good_enzyme_for_high-fidelity_long-range_PCR

I agree with the people above Takara Bio LA is giving good results.

you could improve the isolation if you use Bac clones, if available in your research.

Expand 20 kbPLUS PCR System from Roche

Hi,

TAKARA PrimeSTAR GXL can amplify up to 30 Kb from human genome with a fidelity 10X higher than the standard Taq. I amplified a product of 24 Kb with this enzyme.

For the people using TAKARA LA. Did you use it with human genomic DNA? Is there a possibility to show us the data? How big is your fragment?

I used the TAKARA PrimeSTAR GXL but gave me faint bands with some products that stuck to the wells? Do you have the same issue?

Please see the attached photo.

Thanks.

PRIMESTAR HS DNA Polymerase worth trying

Long range PCR using Takara enzymes is best available solution.