KAPA Real-Time Library Amplification Kits
A novel library amplification solution for optimizing PCR cycle number and reducing PCR-induced bias.
High fidelity PCR is used to selectively enrich library fragments carrying appropriate adaptor sequences and to amplify the amount of DNA prior to sequencing. During PCR enrichment of libraries, minimizing amplification bias is critical to ensure uniform coverage. Introduction of amplification bias requires more total sequencing in order to obtain sufficient coverage in regions of the genome that are underrepresented, increasing sequencing costs. Amplification bias occurs when a DNA polymerase is unable to amplify all targets within a complex population of library DNA with equal efficiency. Bias is further exacerbated when libraries are over-amplified.
KAPA Real-Time PCR Library Amplification Kits are designed to address both sources of PCR-induced bias. The novel KAPA HiFi DNA Polymerase, engineered for high fidelity and processivity, is capable of balanced amplification of complex library DNA. Real-time monitoring of library amplification provides additional information required to minimize over-amplification. Benefits of performing high fidelity, real-time PCR for next-generation sequencing library amplification include:
- Real-time monitoring of amplification allows precise control over the optimal number of PCR cycles, reducing the uncertainty of when to terminate enrichment PCR.
- Real-time amplification workflows are amenable to automation because post-PCR DNA electrophoresis is not required.
- Real-time amplification plots provide quality metrics for individual enriched libraries, eliminating expensive and time-consuming post-enrichment gel electrophoresis.
- Real-time library amplification integrates seamlessly with qPCR-based KAPA Library Quantification Kits.