High sensitivity detection and quantitation of DNA copy number and single nucleotide variants with single color droplet digital PCR.

ABSTRACT: In this study, we present a highly customizable method for quantifying copy number and point mutations utilizing a single-color, droplet digital PCR platform. Droplet digital polymerase chain reaction (ddPCR) is rapidly replacing real-time quantitative PCR (qRT-PCR) as an efficient method of independent DNA quantification. Compared to quantative PCR, ddPCR eliminates the needs for traditional standards; instead, it measures target and reference DNA within the same well. The applications for ddPCR are widespread including targeted quantitation of genetic aberrations, which is commonly achieved with a two-color fluorescent oligonucleotide probe (TaqMan) design. However, the overall cost and need for optimization can be greatly reduced with an alternative method of distinguishing between targ

SUBMITTER: Miotke L 

PROVIDER: S-EPMC3982983 | biostudies-literature | 2014 Mar

REPOSITORIES: biostudies-literature