Genotyping by Fluidigm BioMarkTM IFC Dynamic ArrayTM quantitative PCR.(A) WGA-FLDMqPCR: WGA-amplified DNA and qPCR. Left: double-strand DNA (dsDNA) accumulation curve as a function of the number of cycles. Right: melting curve of the final product. Green: positive control (duck DNA). Red: a hybrid which was positive (containing duck DNA corresponding to the marker tested). Blue: a negative hybrid. Yellow: negative control (hamster DNA). (B) Pre-ampFLDMqPCR: non-amplified DNA, a pre-amplification step with a mix of the 96 primer pairs for the 96 markers tested in the Fluidigm BioMarkTM assay and qPCR. The same markers and controls are used as in (A). The sensitivity is higher in (B), with a lower number of cycles necessary for detection of duck DNA. The negative control and the hybrid not containing duck DNA amplify at a much higher number of cycles and the non-specific products amplified can easily be distinguished by their different melting temperature values (right). In both experiments, no amplification was obtained from water (data not shown).
Rao et al. BMC Genomics 2012 13:513 doi:10.1186/1471-2164-13-513