KAPA HiFi HotStart Ready Mix
KAPA HiFi HotStart DNA Polymerase is a novel B-family DNA polymerase, engineered to have increased affinity for DNA without accessing accessory proteins or binding domains.
The intrinsic high processivity of the enzyme results in significant improvement in yield, speed, and sensitivity when compared with wild-type B-family DNA polymerases. In addition, the ability to amplify long targets, as well as GC- and AT-rich targets, is significantly improved. The enzyme is combined with a proprietary antibody that inactivates the enzyme until the first denaturation step. This prevents nonspecific amplification during reaction setup, increases sensitivity, and improves reaction efficiency.
In the ReadyMix PCR Kit, the enzyme is supplied in a convenient 2X ReadyMix format, containing all reaction components except primers and template. The ReadyMix contains KAPA HiFi HotStart DNA Polymerase (0.5 U per 25 µL reaction) in a proprietary reaction buffer containing dNTPs (0.3 mM of each dNTP at 1X), MgCl2 (2.5 mM at 1X) and stabilizers.
Achieve the highest fidelity
- Increased processivity, strong proofreading activity, and optimized buffer system result in superior accuracy.
- The error rate of KAPA HiFi is 100X lower than Taq, 40X lower than polymerase blends, 3X lower than Pfu Ultra and 2X lower than Phusion.
Improve performance on GC- and AT-rich templates
- Achieve higher success rates across targets up • to 84% GC content.
- Higher yields when amplifying AT-rich templates.
Amplify longer targets with greater sensitivity
- High-fidelity PCR of long and complex genomic templates.
- Improved sensitivity.
- High speed allows significantly shorter reaction times for long-range PCR.
KAPA HiFi HotStart is designed for routine, high-fidelity PCR of a wide range of targets and fragment sizes. It offers error rates approximately 100 times lower than wild-type Taq DNA polymerase, and higher success rates and yields than achievable with wild-type B-family (proofreading) DNA polymerases. In addition, it requires significantly shorter reaction times than wild-type B-family DNA polymerases. KAPA HiFi HotStart DNA Polymerase has 5’g3’ polymerase and 3’g5’ exonuclease (proofreading) activity, but no 5’g3’ exonuclease activity.
The strong 3’g5’ exonuclease activity results in superior accuracy during DNA amplification, lending to KAPA HiFi HotStart DNA Polymerase the lowest published error rate of all B-family DNA polymerases (1 error per 3.6 x 106 nucleotides incorporated). This fidelity is approximately 100 times higher than that of wild-type Taq DNA polymerase, and up to 10 times higher than that of other B-family DNA polymerases and polymerase blends.
DNA fragments generated with KAPA HiFi HotStart ReadyMix may be used for routine downstream analysis and applications, including restriction enzyme digestion, cloning and sequencing. PCR products generated with KAPA HiFi HotStart ReadyMix are blunt-ended, but may be 3’-dA-tailed for cloning into TA cloning vectors (see Important Parameters: TA cloning).