KAPA HiFi Uracil+

KAPA HiFi DNA Polymerase provides high fidelity next-generation sequencing library amplification with the highest yield, lowest bias, and most uniform sequence coverage available. These characteristics are retained in KAPA HiFi Uracil+ DNA Polymerase, a modified version of KAPA HiFi engineered to tolerate uracil residues. The new enzyme is particularly well suited for applications employing bisulfite DNA conversion, which typically produces low concentrations of AT-rich DNA.

KAPA HiFi HotStart Uracil+ ReadyMix (2X) is a ready-to-use cocktail containing all components required for PCR, except primers and template.

Applications:

  • Methylation analysis - amplification of bisulfite-converted DNA for sequencing
  • Amplification of damaged DNA samples
  • Prevention of false-positive results due to carryover amplicon contamination
CodeDescriptionKit ContentsQtyUnit Price
KK2801 KAPA HiFi HotStart Uracil+ ReadyMix (1 x 1.25 mL) 50 x 50 µL reactions. Convenient 2X master mix containing KAPA HiFi HotStart Uracil+ DNA Polymerase, KAPA dNTPs, reaction buffer, and Mg2+ at a 1X final conc. of 2.5 mM. Just add template and primers. **login for pricing
KK2802 KAPA HiFi HotStart Uracil+ ReadyMix (1 x 6.25 mL) 250 x 50 µL reactions. Convenient 2X master mix containing KAPA HiFi HotStart Uracil+ DNA Polymerase, KAPA dNTPs, reaction buffer, and Mg2+ at a 1X final conc. of 2.5 mM. Just add template and primers. **login for pricing

Product Description

KAPA HiFi DNA Polymerase is a novel B-family DNA polymerase exhibiting industry-leading performance in comparison with other high-fidelity proofreading DNA polymerases and polymerase blends. KAPA HiFi DNA Polymerase was engineered for increased affinity to DNA, without the need for accessory protein domains. The intrinsic high processivity of the enzyme results in significant improvements in yield, sensitivity, speed, target length and the ability to amplify difficult amplicons. When used for the amplification of next generation sequencing (NGS) libraries, KAPA HiFi 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. We have inactivated the uracil-binding pocket of KAPA HiFi DNA Polymerase to enable applications that require the amplification of uracil containing DNA, such as bisulfite-sequencing. This new enzyme, KAPA HiFi Uracil+ DNA Polymerase, shows the same high yield, low GC bias and coverage uniformity as the unmodified enzyme. These characteristics are 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’-3’ polymerase and 3’-5’ exonuclease (proofreading) activities, but no 5’-3’ exonuclease activity. The strong 3’-5’ exonuclease activity results in superior accuracy during DNA amplification. A proprietary antibody inactivates the polymerase until the first cycle of thermal denaturation, minimizes spurious amplification products that may result from non-specific priming events during reaction setup and initiation, and increases overall reaction efficiency.

KAPA HiFi HotStart Uracil+ ReadyMix (2X) is a ready-to-use cocktail containing all components required for PCR, except primers and template.

Product Applications

Methylation analysis – amplification of bisulfite- converted DNA for sequencing Sodium bisulfite treatment converts unmethylated cytosines to uracil, while methylated cytosines remain unconverted. Uracil is replaced by thymine during subsequent PCR amplification, allowing base-pair resolution of DNA methylation via sequencing. KAPA HiFi HotStart Uracil+ ReadyMix provides high yields and robust amplification of bisulfite-converted DNA. KAPA HiFi HotStart Uracil+ ReadyMix is especially well suited for NGS applications, providing reduced amplification bias and increased amplification efficiency of bisulfite-converted libraries. Library amplification with KAPA HiFi HotStart Uracil+ provides dramatic improvements in coverage depth uniformity, and more complete representation across reference sequences.

Amplification of damaged DNA samples Cytosine deamination occurs spontaneously over long periods of time, and more rapidly at elevated temperatures, and results in the accumulation of uracil in DNA and among free nucleotides. When other proofreading enzymes fail, KAPA HiFi Uracil+ DNA Polymerase may allow high-fidelity amplification from damaged DNA templates containing uracil.

Prevention of false-positive results due to carryover amplicon contamination KAPA HiFi Uracil+ DNA Polymerase readily incorporates dUTP during amplification, and can therefore be used in conjunction with uracil-DNA-glycosylase (UDG) to prevent carryover contamination. dUTP is added to PCRs so that amplicons that may contaminate subsequent reactions are removed by digestion with UDG prior to amplification.

DocumentTypeDownload
KAPA HiFi HotStart Uracil ReadyMix MSDS Material Safety Data Sheet Download
KAPA HiFi HotStart Uracil ReadyMix TDS Technical Data Sheet Download