KAPA Library Quantification Kit

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Technical Datasheet: KAPA Lib Quant ILMN_9.17 IfU_1 Tags: , , , ,

Description

KAPA Library Quantification Kits contain all the reagents needed for the accurate, reliable and reproducible qPCR-based quantification of next-generation sequencing (NGS) libraries prepared for sequencing on Illumina and IonTorrent platforms. Kits include KAPA SYBR FAST qPCR Master Mix (formulated with different passive reference dyes for different qPCR instruments), a platform-specific library quantification primer premix, and a pre-diluted set of DNA standards. Refer to the Instrument Compatibility Chart for guidance on compatible platforms.

Benefits of KAPA Library Quantification Kits

  • Reliable and sensitive quantification of all sequencing-competent library molecules
  • Accurate and reproducible quantitation across a wide range of library types, concentrations, fragment length distributions and GC content
  • Accurate, equimolar pooling for multiplexed sequencing
  • Flexibility to support manual and automated high-throughput pipelines as well as PCR-free workflows

What makes KAPA Library Quantification Kits highly efficient for library quantification?

KAPA Library Quantification Kits contain KAPA SYBR FAST DNA Polymerase, which was engineered through our directed evolution technology to amplify diverse DNA fragments with similar efficiency. This is important for the amplification of heterogeneous populations such as NGS libraries. Library quantification kits containing wild-type DNA polymerases only count “easy” library molecules.*

View data showing efficient amplification and quantification of diverse libraries by KAPA SYBR FAST DNA Polymerase compared to other options.

How do KAPA Library Quantification Kits Work?

Generation of standard curve and quantification of library concentration

Six pre-diluted DNA Standards and appropriately diluted NGS libraries are amplified using platform-specific qPCRprimers that target adapter sequences. The average Cq value for each DNA Standard is plotted against its known concentration to generate a standard curve. The standard curve is used to convert the average Cq values for diluted libraries to concentration, from which the working concentration of each library is calculated.

Product Highlights

The complete library solution

  • All reagents needed for absolute, qPCR-based quantification of individual NGS libraries or indexed library pools
  • Standard curves to support all library construction workflows, including PCR-free methods
  • Quantification of only/all sequencing-competent library fragments
  • Data analysis templates and assistance available from sequencing.roche.com/support

High consistency and product quality with a proven track record

  • Use of single DNA standard for all library types
  •  Pre-diluted standards with very high lot-to-lot consistency

View data showing high consistency in quantification across library types and consistent amplification efficiency by KAPA Library Quantification Kits.

Reproducible cluster density from samples of variable quality

  • Optimal and predictable cluster densities to maximize sequencing capacity and throughput
  • Reliable results for all library types, including from low-quality FFPE DNA

Equimolar pooling for multiplexed sequencing

  • Equimolar pooling of indexed libraries, irrespective of pre-pooling library concentrations
  • Uniform distribution of reads across all the libraries in a pool

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