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dc.contributor.authorWiseman, Daniel H
dc.contributor.authorSomervaille, Tim C P
dc.date.accessioned2017-08-07T09:58:11Z
dc.date.available2017-08-07T09:58:11Z
dc.date.issued2017
dc.identifier.citationNanofluidic allele-specific digital PCR method for quantifying IDH1 and IDH2 mutation burden in acute myeloid leukemia. 2017, 1633:235-255 Methods Mol. Biol.en
dc.identifier.issn1940-6029
dc.identifier.pmid28735491
dc.identifier.doi10.1007/978-1-4939-7142-8_15
dc.identifier.urihttp://hdl.handle.net/10541/620479
dc.description.abstractPrecise quantitation of allelic burden for a pathogenic mutation has diverse clinical and research applications but can be difficult to achieve with conventional qPCR-based techniques, especially at lower mutant allele frequencies. Digital PCR overcomes many of the limitations of qPCR and can be highly quantitative even for single-nucleotide variants, with distinct advantages over next-generation sequencing approaches. Here we describe a method combining the principles of TaqMan(®)-chemistry SNP genotyping with microfluidic digital PCR to generate a highly sensitive, quantitative allele-specific digital PCR assay for the six most common IDH1 and IDH2 mutations encountered in myeloid malignancy. The concept and approach could easily be applied to other suitable SNVs.
dc.language.isoenen
dc.rightsArchived with thanks to Methods in molecular biology (Clifton, N.J.)en
dc.titleNanofluidic allele-specific digital PCR method for quantifying IDH1 and IDH2 mutation burden in acute myeloid leukemia.en
dc.typeArticleen
dc.contributor.departmentLeukemia Biology Laboratory, Cancer Research UK Manchester Institute, The University of Manchester, Manchester, M20 4BX, UKen
dc.identifier.journalMethods in Molecular Biologyen
html.description.abstractPrecise quantitation of allelic burden for a pathogenic mutation has diverse clinical and research applications but can be difficult to achieve with conventional qPCR-based techniques, especially at lower mutant allele frequencies. Digital PCR overcomes many of the limitations of qPCR and can be highly quantitative even for single-nucleotide variants, with distinct advantages over next-generation sequencing approaches. Here we describe a method combining the principles of TaqMan(®)-chemistry SNP genotyping with microfluidic digital PCR to generate a highly sensitive, quantitative allele-specific digital PCR assay for the six most common IDH1 and IDH2 mutations encountered in myeloid malignancy. The concept and approach could easily be applied to other suitable SNVs.


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