Next Generation Sequencing with a High Performance qPCR Solution

The new KAPA Library Quantification Kits available from Anachem utilise the power of Quantitative PCR (qPCR) to provide users with an accurate and sensitive method for quantifying Next Generation Sequencing (NGS) libraries. Standard methods for quantifying NGS libraries have a variety of disadvantages. Electrophoresis and spectrophotometry measure total nucleic acid concentrations, whereas optimal cluster density or template-to-bead ratio depend on the appropriate concentration of PCR-amplifiable DNA molecules. These methods also have low sensitivity, consuming nanograms of precious samples and are not suitable for high-throughput workflows.

qPCR, however, is a highly sensitive approach that uses a minimal amount of material and eliminates the need for time-consuming and expensive titrations. It specifically quantifies only PCR-competent DNA molecules and due to its high sensitivity, allows for the accurate quantification of low concentration libraries and significantly reduces cluster density variations. It can also be automated for highthroughput applications.

The Broad Institute, Cambridge, MA USA stated: “Before qPCR was adopted for library quantification, cluster density was extremely variable. Implementation of the KAPA Library Quantification Kit into our sequencing workflows resulted in a significant reduction in variability across multiple libraries, negating the need for cluster amplification titration runs."

The KAPA Library Quantification kits are optimised for the Illumina Genome Analyser, Roche 454 Titanium series, and Roche 454 FLX series platforms and include defined, reliable DNA concentration standards and state-of-the-art qPCR reagents, containing a DNA polymerase engineered for SYBR® Green-based qPCR through a process of molecular evolution to provide a rapid, sensitive, and reliable method for quantifying PCR-amplifiable molecules in next-generation sequencing DNA libraries.