Quantitative DNA Analysis Using Droplet Digital PCR.
Droplet digital PCR (ddPCR) is based on the isolated amplification of thousands of individual DNA molecules simultaneously, with each molecule compartmentalized in a droplet. The presence of amplified product in each droplet is indicated by a fluorescent signal, and the proportion of positive droplets allows the precise quantification of a given sequence. In this chapter we briefly outline the basis of ddPCR, and describe two different applications using the Bio-Rad QX200 system: genotyping copy number variation and quantification of Illumina sequencing libraries.
Leiden Genome Technology Center, Department of Human Genetics, Leiden University Medical Center, Einthovenweg 20, 2333 ZC, Leiden, The Netherlands.
Rolf H A M
RH
Vossen
Leiden Genome Technology Center, Department of Human Genetics, Leiden University Medical Center, Einthovenweg 20, 2333 ZC, Leiden, The Netherlands. s.j.white@lumc.nl.
Stefan J
SJ
White
D004247:DNA / Q000235:genetics*; D056915:DNA Copy Number Variations; D059014:High-Throughput Nucleotide Sequencing; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods*; D060888:Real-Time Polymerase Chain Reaction
D004247:DNA
Copy number variation; DNA quantitation; Digital PCR; NGS