Open AccessDigital Droplet PCR: CNV Analysis and Other Applications
Author(s) -
Mazaika Erica,
Homsy Jason
Publication year - 2014
Publication title -
current protocols in human genetics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.282
H-Index - 30
eISSN - 1934-8258
pISSN - 1934-8266
DOI - 10.1002/0471142905.hg0724s82
Subject(s) - digital polymerase chain reaction , computational biology , computer science , biology , polymerase chain reaction , genetics , gene
Digital droplet PCR (ddPCR) is an assay that combines state‐of‐the‐art microfluidics technology with TaqMan‐based PCR to achieve precise target DNA quantification at high levels of sensitivity and specificity. Because quantification is achieved without the need for standard assays in an easy to interpret, unambiguous digital readout, ddPCR is far simpler, faster, and less error prone than real‐time qPCR. The basic protocol can be modified with minor adjustments to suit a wide range of applications, such as CNV analysis, rare variant detection, SNP genotyping, and transcript quantification. This unit describes the ddPCR workflow in detail for the Bio‐Rad QX100 system, but the theory and data interpretation are generalizable to any ddPCR system. Curr. Protoc. Hum. Genet . 82:7.24.1‐7.24.13. © 2014 by John Wiley & Sons, Inc.