Abstract
Digital PCR is a valuable tool to quantify next-generation sequencing (NGS) libraries precisely and accurately. Accurately quantifying NGS libraries enable accurate loading of the libraries on to the sequencer and thus improve sequencing performance by reducing under and overloading error. Accurate quantification also benefits users by enabling uniform loading of indexed/barcoded libraries which in turn greatly improves sequencing uniformity of the indexed/barcoded samples. The advantages gained by employing the Droplet Digital PCR (ddPCR™) library QC assay includes the precise and accurate quantification in addition to size quality assessment, enabling users to QC their sequencing libraries with confidence.
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Further Reading
Laurie MT, Bertout JA, Taylor SD, Burton JN, Shendure JA, Bielas JH (2013) Simultaneous digital quantification and fluorescence-based size characterization of massively parallel sequencing libraries. BioTechniques 55(2):61–67
Buehler B, Hogrefe HH, Scott G, Ravi H, Pabón-Peña C, O’Brien S, Formosa R, Happe S (2010) Rapid quantification of DNA libraries for next-generation sequencing. Methods 50:S15–S18
QPCR NGS Library Quantification Kit (illumina GA), Part Number G4880A, Publication number 5990–3065. (2012). Agilent
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Sequencing Library qPCR Quantification Guide Catalog # SY-930-1010, Part #11322363 Rev. C (2011). Illumina
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Heredia, N.J. (2018). Droplet Digital™ PCR Next-Generation Sequencing Library QC Assay. In: Karlin-Neumann, G., Bizouarn, F. (eds) Digital PCR. Methods in Molecular Biology, vol 1768. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7778-9_27
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DOI: https://doi.org/10.1007/978-1-4939-7778-9_27
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Publisher Name: Humana Press, New York, NY
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Online ISBN: 978-1-4939-7778-9
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