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dc.contributor.authorPender, Aen_US
dc.contributor.authorGarcia-Murillas, Ien_US
dc.contributor.authorRana, Sen_US
dc.contributor.authorCutts, RJen_US
dc.contributor.authorKelly, Gen_US
dc.contributor.authorFenwick, Ken_US
dc.contributor.authorKozarewa, Ien_US
dc.contributor.authorGonzalez de Castro, Den_US
dc.contributor.authorBhosle, Jen_US
dc.contributor.authorO'Brien, Men_US
dc.contributor.authorTurner, NCen_US
dc.contributor.authorPopat, Sen_US
dc.contributor.authorDownward, Jen_US
dc.date.accessioned2018-11-29T09:20:04Z
dc.date.issued2015-01en_US
dc.identifier.citationPloS one, 2015, 10 (9), pp. e0139074 - ?en_US
dc.identifier.urihttps://repository.icr.ac.uk/handle/internal/2962
dc.identifier.eissn1932-6203en_US
dc.identifier.doi10.1371/journal.pone.0139074en_US
dc.description.abstractDroplet digital PCR (ddPCR) can be used to detect low frequency mutations in oncogene-driven lung cancer. The range of KRAS point mutations observed in NSCLC necessitates a multiplex approach to efficient mutation detection in circulating DNA. Here we report the design and optimisation of three discriminatory ddPCR multiplex assays investigating nine different KRAS mutations using PrimePCR™ ddPCR™ Mutation Assays and the Bio-Rad QX100 system. Together these mutations account for 95% of the nucleotide changes found in KRAS in human cancer. Multiplex reactions were optimised on genomic DNA extracted from KRAS mutant cell lines and tested on DNA extracted from fixed tumour tissue from a cohort of lung cancer patients without prior knowledge of the specific KRAS genotype. The multiplex ddPCR assays had a limit of detection of better than 1 mutant KRAS molecule in 2,000 wild-type KRAS molecules, which compared favourably with a limit of detection of 1 in 50 for next generation sequencing and 1 in 10 for Sanger sequencing. Multiplex ddPCR assays thus provide a highly efficient methodology to identify KRAS mutations in lung adenocarcinoma.en_US
dc.formatElectronic-eCollectionen_US
dc.format.extente0139074 - ?en_US
dc.languageengen_US
dc.language.isoengen_US
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/en_US
dc.subjectCell Line, Tumoren_US
dc.subjectClone Cellsen_US
dc.subjectHumansen_US
dc.subjectCarcinoma, Non-Small-Cell Lungen_US
dc.subjectLung Neoplasmsen_US
dc.subjectFormaldehydeen_US
dc.subjectDNA, Neoplasmen_US
dc.subjectParaffin Embeddingen_US
dc.subjectTissue Fixationen_US
dc.subjectTemperatureen_US
dc.subjectGene Frequencyen_US
dc.subjectMutationen_US
dc.subjectProto-Oncogene Proteins p21(ras)en_US
dc.subjectHigh-Throughput Nucleotide Sequencingen_US
dc.subjectGenotyping Techniquesen_US
dc.subjectMultiplex Polymerase Chain Reactionen_US
dc.titleEfficient Genotyping of KRAS Mutant Non-Small Cell Lung Cancer Using a Multiplexed Droplet Digital PCR Approach.en_US
dc.typeJournal Article
dcterms.dateAccepted2015-09-09en_US
rioxxterms.versionofrecord10.1371/journal.pone.0139074en_US
rioxxterms.licenseref.startdate2015-01en_US
rioxxterms.typeJournal Article/Reviewen_US
dc.relation.isPartOfPloS oneen_US
pubs.issue9en_US
pubs.notesNot knownen_US
pubs.organisational-group/ICR
pubs.organisational-group/ICR/Primary Group
pubs.organisational-group/ICR/Primary Group/ICR Divisions
pubs.organisational-group/ICR/Primary Group/ICR Divisions/Breast Cancer Research
pubs.organisational-group/ICR/Primary Group/ICR Divisions/Breast Cancer Research/Molecular Oncology
pubs.organisational-group/ICR/Primary Group/ICR Divisions/Cancer Biology
pubs.organisational-group/ICR/Primary Group/ICR Divisions/Cancer Biology/Lung Cancer Group
pubs.organisational-group/ICR/Primary Group/ICR Divisions/Clinical Studies
pubs.organisational-group/ICR/Primary Group/ICR Divisions/Clinical Studies/Thoracic Oncology
pubs.organisational-group/ICR/Primary Group/ICR Divisions/Clinical Studies/Thoracic Oncology/Thoracic Oncology (hon.)
pubs.organisational-group/ICR/Primary Group/ICR Divisions/Clinical Studies/Treatment of thoracic tumours
pubs.organisational-group/ICR/Primary Group/ICR Divisions/Clinical Studies/Treatment of thoracic tumours/Treatment of thoracic tumours (hon.)
pubs.organisational-group/ICR/Primary Group/ICR Divisions/Molecular Pathology
pubs.organisational-group/ICR/Primary Group/ICR Divisions/Molecular Pathology/Lung Cancer Group
pubs.organisational-group/ICR/Primary Group/Royal Marsden Clinical Units
pubs.volume10en_US
pubs.embargo.termsNot knownen_US
icr.researchteamMolecular Oncologyen_US
icr.researchteamThoracic Oncologyen_US
icr.researchteamTreatment of thoracic tumoursen_US
icr.researchteamLung Cancer Groupen_US
dc.contributor.icrauthorGarcia-Murillas, Isaacen_US
dc.contributor.icrauthorTurner, Nicholasen_US
dc.contributor.icrauthorDownward, Julian David Harryen_US
dc.contributor.icrauthorGonzalez de Castro, Daviden_US
dc.contributor.icrauthorO'Brien, Maryen_US
dc.contributor.icrauthorPender, Alexandraen_US
dc.contributor.icrauthorPopat, Sanjayen_US


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Except where otherwise noted, this item's license is described as http://creativecommons.org/licenses/by/4.0/