Optimization of digital droplet polymerase chain reaction for quantification of genetically modified organisms.

ABSTRACT: Digital PCR in droplets (ddPCR) is an emerging method for more and more applications in DNA (and RNA) analysis. Special requirements when establishing ddPCR for analysis of genetically modified organisms (GMO) in a laboratory include the choice between validated official qPCR methods and the optimization of these assays for a ddPCR format. Differentiation between droplets with positive reaction and negative droplets, that is setting of an appropriate threshold, can be crucial for a correct measurement. This holds true in particular when independent transgene and plant-specific reference gene copy numbers have to be combined to determine the content of GM material in a sample. Droplets which show fluorescent units ranging between those of explicit positive and negative droplets are called '

SUBMITTER: Gerdes L 

PROVIDER: S-EPMC4827695 | biostudies-literature | 2016 Mar

REPOSITORIES: biostudies-literature