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Rare Variant Quantitation Using Droplet Digital PCR

  • Brian Parkin
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1881)

Abstract

Droplet digital PCR (ddPCR) allows precise quantitation of individual copies of DNA. Using allele-specific fluorescent oligonucleotide probes, ddPCR can enumerate both mutant and wild-type alleles enabling highly sensitive detection and quantitation of the variant allele frequency of mutated genes. In this protocol, we describe a method for using ddPCR to detect mutations in genomic DNA with a sensitivity of up to 1 in 50,000 DNA copies.

Key words

Digital PCR Rare variant detection Mutation quantitation Polymerase chain reaction Allele-specific probes Locked nucleic acid 

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Division of Hematology and Oncology, Department of Internal MedicineUniversity of MichiganAnn ArborUSA
  2. 2.Ann Arbor VA Healthcare SystemAnn ArborUSA

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