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For Research Use Only. Not for use in diagnostic procedures. Others KAPA Library Quantification Primer Premix and KAPA SYBR FAST RUO KAPA Library Quantification Primer Premix and KAPA SYBR FAST SEQ-KAPA-0137 07 960 441 001 7 960 441 001 07960441001 7960441001 07960441001 LQ Primer+KSF (Uni) LQ Primer+KSF (Uni) 07613336112946 Reagents, kits KK4923 500 reactions Not Available 07 960 573 001 7 960 573 001 07960573001 7960573001 07960573001 LQ Primer+KSF (LC480) LQ Primer+KSF (LC480) 07613336113066 Reagents, kits KK4953 500 reactions Not Available KAPA Library Quantification Kits provide all the reagents needed for absolute, qPCR-based quantification of Illumina libraries flanked by the P5 and P7 flow cell oligo sequences. Kits contain:
Library Quantification DNA Standards 1 – 6 (a 10-fold dilution series of a linear, 452 bp template)
Library Quantification Primer Premix (10X), containing the following primers:
Primer 1: 5'-AAT GAT ACG GCG ACC ACC GA-3'
Primer 2: 5'-CAA GCA GAA GAC GGC ATA CGA-3'
KAPA SYBR® FAST qPCR Master Mix (2X), available with various passive reference dyes (Table 1).
Library quantification is performed by amplifying the set of six pre-diluted DNA Standards and diluted library samples by qPCR, using the KAPA SYBR FAST qPCR Master Mix and primers targeting the Illumina P5 and P7 flow cell oligo sequences. The average Cq score for each DNA Standard is plotted against log10 (concentration in pM) to generate a standard curve. The concentrations of diluted library samples are then calculated against the standard curve, using absolute quantification.
KAPA Library Quantification Kits are rigorously tested to ensure minimal lot-to-lot variation. The kit contains the novel KAPA SYBR FAST DNA Polymerase, engineered through a process of directed evolution for high- performance SYBR Green I-based qPCR. The ability of the engineered polymerase to amplify diverse DNA fragments with similar efficiency enables the use of a universal standard for the reliable quantification of all Illumina libraries with an average fragment length of up to 1 kb, irrespective of library type or GC content.
KAPA SYBR FAST is an antibody-mediated hot start DNA polymerase formulation. KAPA Library Quantification Kits are therefore suitable for use with automated liquid handling systems for high-throughput sample quantification. en KAPA Library Quantification Kits are designed for the accurate and reproducible quantification of libraries prepared for Illumina sequencing. Any library with a concentration >0.0002pM that contains sequences complementary to the primers in the Primer Premix (10X) can be quantified with the kit, irrespective of the library type, how it was constructed, or on which Illumina instrument it will be sequenced.
The kit supports quantification of libraries across a range of GC contents and average fragment lengths up to 1 kb.
In addition to NGS library quantification, the kit can also be used to detect library contamination in work spaces used during the preparation of Illumina libraries.
The KAPA Library Quantification assay consists of repetitive pipetting steps, which can easily be automated. The use of an automated liquid handling system is highly recommended for high-throughput NGS pipelines. Please refer to the KAPA Library Quantification Technical Guide for more information. en