KAPA Library Quantification Kits

For Research Use Only. Not for use in diagnostic procedures.

KAPA Library Quantification Kits


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. 

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.

data showing efficient amplification and quantification of diverse libraries

Figure 1. KAPA SYBR FAST exhibits high-efficiency amplification and library quantification. Amplification efficiencies achieved with KAPA SYBR FAST for ten diverse amplicons, plotted against GC content. All ten amplicons were amplified with similar efficiency (within the optimal range of 95% –105%). This confirms that KAPA SYBR FAST is the ideal enzyme for the amplification of heterogenous populations of DNA fragments, such as NGS libraries.

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 qPCR primers 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.


Figure 3 Six pre-diluted DNA Standards

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
Figure 3 KAPA SYBR FAST vs. 2 competitors

Figure 3. DNA fragment size distributions for an NGS library before (grey fill) and after qPCR amplification with KAPA SYBR FAST and two competitor kits containing wild-type Taq polymerase. Reactions were performed with the following cycling protocol: 95°C for 10 min, followed by 40 cycles of 95°C for 10 sec, and 60°C for 45 sec. The KAPA SYBR FAST quantification product is representative of the template DNA (NGS library), whereas the quantification products generated with wild-type enzymes are not.

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
Figure 8 Cluster densities from samples
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

View data showing uniform distribution of reads in multiplexed sequencing pools using KAPA Library Quantification Kits.


  • Compatible Platform

    Illumina HiSeq, NextSeq, MiSeq, and GAIIx Ion Torrent Proton and PGM 454 GS FLX+ and GS Junior SOLiD 5500 series

  • Library Type


  • Starting Material

    Any NGS library ready for cluster amplification or emPCR

  • Standard curve concentration range

    20–0.0002 pM for Illumina libraries 83–0.00083 pM for Ion Torrent and 454 libraries 10–0.0001 pg/µL for SOLiD libraries

  • Sequencing Applications

    Whole Genome Sequencing, Whole Exome Sequencing, Targeted Sequencing (custom panels), RNA-Seq, ChIP-Seq, Amplicon Sequencing, Methyl-Seq

Kits can be stored at -20˚C.

Complete kits include KAPA SYBR FAST qPCR Master Mix (2X), Primer Premix (10X) and a set of 6 DNA Standards. Reagents for qPCR (KAPA SYBR FAST qPCR Master Mix and Primer Premix) and DNA Standards (1–6) are also sold separately. Where noted, Master Mixes contain instrument-specific reference dyes, while the Universal kit includes ROX High and ROX Low (both 50X) separately.

Components–Illumina Platform

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Components–Illumina Platform

  Kit Code 10X Illumina Primer Premix 2X KAPA SYBR FAST qPCR Master Mix (Universal) 50X KAPA SYBR FAST ROX High 50X KAPA SYBR FAST ROX Low 2X KAPA SYBR FAST qPCR Master Mix
(with equipment-specific dye)
qPCR Master Mix
(ROX Low)
Illumina DNA Standards 1-6
Complete Kit (universal)
KK4824 X X X X     X
Complete Kit (with equipment-specific dye)
KK4835 X       X

Complete Kit (ROX Low)
KK4873 X         X X
qPCR MasterMix (universal) and Primer Premix only KK4923 X X X X      

qPCR MasterMix (with equipment-specific  reference dye)

 and Primer Premix only

KK4933 X

qPCR MasterMix (ROX Low) 
and Primer Premix only
KK4973 X         X  

Illumina Platforms

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Illumina Platforms

Roche Cat. No
Kit Size
How to buy
Complete kit (Universal) 500 x 20 µL reactions
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Complete kit (ABI Prism®) 500 x 20 µL reactions
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Complete kit (BioRad® iCycler only)

500 x 20 µL reactions
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Complete kit (optimized for Roche® LightCycler 480) 500 x 20 µL reactions
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Complete Kit (ROX Low) 500 x 20 µL reactions
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qPCR MasterMix (Universal) and Primer Premix only 500 x 20 µL reactions
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qPCR MasterMix (optimized for Roche LightCycler 480) and Primer Premix only 500 x 20 µL reactions
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