KAPA HiFi HotStart provides high fidelity NGS library amplification with the highest yield, lowest bias, and most uniform sequence coverage available. These characteristics are retained in KAPA HiFi HotStart Uracil+, a modified version of KAPA HiFi HotStart, engineered to tolerate uracil residues. KAPA HiFi HotStart Uracil+ ReadyMix contains all components for PCR, except primers and template.
KAPA HiFi HotStart is a novel B-family DNA polymerase, engineered to have increased affinity for DNA, without the need for accessory proteins or DNA-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 and polymerase blends. When used for the amplification of next-generation sequencing (NGS) libraries, KAPA HiFi HotStart DNA Polymerase exhibits high yields with minimal amplification bias and provides extremely uniform sequence coverage.
The read-ahead function of proofreading DNA polymerases detects pro-mutagenic uracil residues in the template strand and prevents further strand extension, thereby reducing or completely inhibiting PCR amplification. In KAPA HiFi HotStart Uracil+ DNA Polymerase, this uracilbinding pocket is inactivated to enable the amplification of uracil-containing DNA. The enzyme shows the same high yield, low GC-bias and coverage uniformity as the unmodified KAPA HiFi HotStart DNA Polymerase, making it particularly advantageous for applications employing bisulfite DNA conversion, which typically produces low concentrations of AT-rich DNA.
KAPA HiFi HotStart Uracil+ 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. A proprietary antibody inactivates the polymerase until the first cycle of denaturation, minimizing spurious amplification products that may result from nonspecific priming events during reaction setup and initiation, and increasing overall reaction efficiency.
KAPA HiFi HotStart Uracil+ ReadyMix (2X) is a ready-touse cocktail containing all components required for PCR, except primers and template. The ReadyMix contains 0.2 mM of each dNTP (dATP, dCTP, dGTP, dTTP), and does not contain dUTP.
The name of this product was recently changed. The old name was: KAPA High Fidelity HotStart Uracil with ReadyMix