![]() (1) MTX Terminator TNE212D 1200 Watt 2 Ohm Dual 12” Subwoofers + Sealed Sub Box.The old product name was KAPA Real-Time Library Amplification Kit.įind here our other Library Amplification solutions. PCR products generated with KAPA HiFi HotStart Real-time PCR Master Mix (2X) are blunt-ended, but may be 3′-dA-tailed for cloning into TA cloning vectors. The presence of SYBR Green I dye in the reaction does not compromise fidelity.ĭNA fragments generated with KAPA HiFi HotStart Real-time PCR Master Mix (2X) may be used for routine downstream applications, including restriction enzyme digestion and sequencing. This fidelity is approximately 100X higher than that of wild-type Taq and up to 30X higher than polymerase blends. The error rate of KAPA HiFi HotStart DNA Polymerase is ~2.8 x 10-7. The strong 3’g5′ exonuclease activity results in superior accuracy during DNA amplification. KAPA HiFi HotStart DNA Polymerase has 5’g3′ polymerase and 3’g5′ exonuclease (proofreading) activities, but no 5’g3′ exonuclease activity. This eliminates spurious amplification products resulting from non-specific priming events during reaction setup and initiation and increases overall reaction efficiency. In the HotStart formulation, a proprietary antibody inactivates the polymerase until the first denaturation step. These enhancements result in lower amplification bias which leads to more uniform sequence coverage. The intrinsic high processivity of the enzyme results in significant improvements in yield, sensitivity, speed, target length, and the ability to amplify difficult amplicons. KAPA HiFi DNA Polymerase was engineered for increased affinity to DNA, without the need for accessory protein domains. It is an antibody-based hot start formulation of KAPA HiFi DNA Polymerase, a novel B-family DNA polymerase exhibiting industry leading performance in comparison with other high-fidelity (B-family) DNA polymerases and polymerase blends. Seamless integration with KAPA Library Quantification Kits.Real-time amplification plots provide quality metrics for individual enriched libraries, eliminating expensive and time-consuming post-enrichment gel electrophoresis and identifying inconsistencies in library preparation.Real-time amplification workflows are amenable to automation.Real-time monitoring of amplification allows precise control over the optimal number of PCR cycles.Benefits of performing high fidelity, real-time PCR for next-generation sequencing library amplification include: Real-time monitoring of library amplification provides additional information required to minimize over-amplification. The novel KAPA HiFi DNA Polymerase, engineered for high fidelity and processivity, is capable of balanced amplification of complex library DNA. KAPA Real-time Library Amplification Kits are designed to address both sources of PCR-induced bias. Bias is further exacerbated when libraries are over-amplified. Amplification bias occurs when a DNA polymerase is unable to amplify all targets within a complex population of library DNA with equal efficiency. During PCR enrichment of libraries, minimizing amplification bias is critical to ensure uniform sequence coverage. High-fidelity PCR is used to selectively enrich library fragments carrying appropriate adapter sequences and to amplify the amount of DNA prior to sequencing. Four fluorescent standards are supplied and are used to define a window for optimal amplification The master mix contains KAPA HiFi HotStart DNA Polymerase in a proprietary reaction buffer, dNTPs, MgCl2 (2.5 mM at 1X), SYBR® Green I dye, and stabilizers. KAPA HiFi HotStart Real-Time Library Amplification Kits contain KAPA HiFi HotStart Real-Time PCR Master Mix (2X), a ready-to-use cocktail containing all componentsįor PCR, except primers and template. KAPA HiFi HotStart Real-Time Library Amplification Kits are designed for the amplification of NGS libraries prepared for Illumina sequencing. KAPA HiFi HotStart Real-Time Library Amp Kit
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