Kapa biosystems/KAPA Library Quantification Kits/KK4838/500 x 20 µL reactions

Description:

Complete kit (ABI Prism®)

KAPA Library Quantification Kits

for Next-Generation Sequencing

Our standards don’t change. Neither should yours.

KAPA Library Quantification Kits contain all the reagents needed for the accurate, reliable and reproducible qPCR-based quantification of NGS libraries prior to pooling for capture or flow cell amplification. Kits contain KAPA SYBR^® FAST qPCR Master Mix, optimized for high-performance SYBR Green I-based qPCR. The engineered KAPA SYBR FAST DNA Polymerase contained in the Master Mix amplifies GC- and AT-rich DNA fragments of different lengths with similar efficiency, making it the only qPCR Master Mix capable of accurate qPCR-based quantification of NGS libraries.* The pre-diluted set of DNA Standards contained in each kit is carefully quality-controlled to ensure lot-to-lot consistency and avoid data drift over time. Optimized Primer Premixes ensure optimal PCR efficiency.

Kits with DNA Standards and Primer Premixes for Illumina^® and Ion Torrent™ sequencers are available. Please reference the Instrument Compatibility Chart for guidance on compatible platforms.

NEW! Improve experimental designs with our Technical Guide for Illumina Platforms

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

Product Highlights

qPCR-based KAPA Library Quantification Kits reduce variability in cluster density compared to Agilent Bioanalyzer. Human exome samples were prepared using Nimblegen target capture. All preparations were performed in a 96-well plate format on a liquid handling system. Average number of clusters per tile are shown for paired-end, 76-bp Illumina GAIIx runs. Data courtesy of the University of Washington. Data on file.

Compared to spectrophotometric, fluorimetric, and electrophoretic methods, qPCR only quantifies adaptor-ligated and amplifiable library molecules. The ligation of adaptor sequences (pink and blue) to library DNA molecules (grey) results in a mixed population of molecules without the correct adaptor configuration. Electrophoresis and spectrophotometry measure total nucleic acid concentrations, whereas optimal cluster density or template-to-bead ratio depends on the appropriate concentration of PCR-amplifiable DNA molecules. Only qPCR-based library quantification methods count library molecules with the correct adaptor sequence configuration. Data on file.

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Accurately quantify PCR-competent sequencing templates

qPCR only “counts” those library molecules that can be sequenced
More consistent cluster densities or template-to-bead ratios enable optimal utilization of sequencing resources

Equimolar pooling of indexed libraries. Eleven indexed Illumina TruSeq libraries were quantified by qPCR using the KAPA Library Quantification Kit, and then combined to achieve equal final concentrations in two separate pools for multiplexed sequencing on different flow-cell lanes. The eleven libraries ranged ~11-fold in concentration from 0.67 pM to 7.65 pM, while representation of each index varied between 90% and 127% of expected assigned reads per lane. Data on file.

Improve pooling for multiplexed sequencing

Library concentrations determined by qPCR are more reliable compared to other methods*
The engineered KAPA SYBR FAST qPCR Master Mix enables accurate pooling of libraries with extreme GC-contents or longer inserts*

Nine human DNA libraries and two microbial DNA libraries (Rhodococcus sp. ~70% GC and Staphylococcus sp. ~35% GC) were used to compare quantification results obtained with distinct lots (”Lot 1” and “Lot 2”), and distinct sets of reagents from the same lot (”Set 1” and “Set 2”) of KAPA Library Quantification Kits for Illumina platforms. Data on file.

Eliminate drift with carefully quality-controlled DNA Standards

Prediluted DNA Standards eliminate quantification errors associated with preparation of standard curves using in-house standards*
KAPA DNA Standards undergo strict quality control to ensure lot-to-lot consistency and eliminate data drift over time

KAPA Library Quantification methods (96- and 384-well) format are available for the Sciclone NGS and NGSx workstation (PerkinElmer) and other instruments used in high-throughput NGS sample preparation pipelines.

Improve throughput with automation

Library quantification assay is compatible with 96- and 384-well format
Library dilution, reaction setup and data analysis can be automated for HTP pipelines
Learn More about KAPA Automated Solutions

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Related Products

Normalizing libraries for pooling and cluster amplification? Check out these Kapa NGS products to improve your workflow and results:

KAPA hgDNA Quantification and QC Kit

KAPA Library Amplification Kits

KAPA Hyper Prep Kits

KAPA Stranded mRNA-Seq Kits

Applications:

Applications

Quantification of libraries prior to pooling for multiplexed sequencing
Quantification of individual libraries or library pools prior to cluster amplification or emPCR
Quantification of libraries prior to and after hybridization capture
Quality control, optimization and troubleshooting of library construction processes or workflows

Kit Specifications and Contents / Storage:

Kit Specifications and Contents / Storage

Kits can be stored for up to 12 months 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

Components–Ion Torrent™ Platform

LQK-Ion_Component Chart

Specifications

Compatible Platforms

Illumina HiSeq, NextSeq, MiSeq, and GAIIxIon Torrent Proton and PGM454 GS FLX+ and GS JuniorSOLiD 5500 series

Library Type

Any

Starting Material

Any NGS library ready for cluster amplification or emPCR

Standard curve concentration range

20–0.0002 pM for Illumina libraries83–0.00083 pM for Ion Torrent and 454 libraries10–0.0001 pg/µL for SOLiD libraries

Sequencing Applications

Whole Genome Sequencing Whole Exome Sequencing Targeted Sequencing (custom panels)RNA-SeqChIP-SeqAmplicon Sequencing Methyl-Seq

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