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Genetics

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JoVE Journal Genetics
Single Droplet Digital Polymerase Chain Reaction for Comprehensive and Simultaneous Detection of Mutations in Hotspot Regions
 

Single Droplet Digital Polymerase Chain Reaction for Comprehensive and Simultaneous Detection of Mutations in Hotspot Regions

Article DOI: 10.3791/58051-v • 08:23 min • September 25th, 2018
September 25th, 2018 •
1,2, 1, 1, 1,3,4, 1,3,5, 1,6, 1
1Circulating Tumor Biomarkers Laboratory, SiRIC, Translational Research Department, Institut Curie, PSL Research University, 2CNRS UMR144, Institut Curie, PSL Research University, 3Department of Medical Oncology, Institut Curie, PSL Research University, 4University Versailles Saint-Quentin-en-Yvelines, 5University Paris Descartes, 6Inserm U830, Institut Curie, PSL Research University

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Here, we present a protocol to accurately quantify multiple genetic alterations of a target region in a single reaction using drop-off ddPCR and a unique pair of hydrolysis probes.

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Keywords: Single Droplet Digital PCR Circulating Tumor DNA Mutation Detection Hotspot Regions Cancer Diagnosis Absolute Quantification Molecular Analysis QPCR Alternative Plasma Extraction Proteinase K Lysis Buffer Silica Membrane Vacuum Pump
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