Analyzing Copy Number Variation with Droplet Digital PCR.

Methods Mol Biol
Authors
Keywords
Abstract

Many genomic segments vary in copy number among individuals of the same species, or between cancer and normal cells within the same person. Correctly measuring this copy number variation is critical for studying its genetic properties, its distribution in populations and its relationship to phenotypes. Droplet digital PCR (ddPCR) enables accurate measurement of copy number by partitioning a PCR reaction into thousands of nanoliter-scale droplets, so that a genomic sequence of interest-whose presence or absence in a droplet is determined by end-point fluorescence-can be digitally counted. Here, we describe how we analyze copy number variants using ddPCR and review the design of effective assays, the performance of ddPCR with those assays, the optimization of reactions, and the interpretation of data.

Year of Publication
2018
Journal
Methods Mol Biol
Volume
1768
Pages
143-160
Date Published
2018
ISSN
1940-6029
DOI
10.1007/978-1-4939-7778-9_9
PubMed ID
29717442
Links
Grant list
R01 HG006855 / HG / NHGRI NIH HHS / United States