Product Catalogue

"Before qPCR was adopted for library quantification, cluster density was extremely variable. Implementation of the KAPA Library Quantification Kit into our sequencing workflows resulted in a significant reduction in variability across multiple libraries, negating the need for cluster amplification titration runs".
- The Broad Institute, Cambridge, MA U.S.A.
 

I have now tried the KAPA HiFi HotStart enzyme and sequenced cloned PCR fragment to see how many introduced PCR mutation this enzyme gives.

Template: cDNA from the C2C12 cell line

Expected product size: 2.3 kb

Reaction setup: As described in the manufacturers manual

Cycles:

95° 3'
98° 20" 60° 15" 72° 90" 40x
72° 3'

Cloned and sequenced three individual clones.

Mutation: 0 (zeeeeero)

I have also used this enzyme for low complexity amplifications (plasmid as template and 16 cycles) and have not seen a single introduced mutation.

KAPA HiFi HotStart is absolutely the best enzyme I have ever used when high fidelity is desired and is my future choice for all my PCR applications, instead of the many other enzymes I have used and tried over nearly two decades in science.

KAPA HiFi HotStart has also, in my hands, proven to be very robust; it has worked every time even with known difficult template and primer pairs.

Göran Hjälm, Assc. Professor
Uppsala University
Sweden