dPCR CNV Probe Gene of Interest Assay

From QIAGEN, this dPCR assay supports copy number analysis applications such as the quantification of gene-specific regions in human, mouse, or rat DNA on the QIAcuity digital PCR instrument. It is intended for use with the QIAcuity Probe PCR Kit or QIAcuity High Multiplex Probe PCR Kit, in singleplex or multiplex reaction formats, to detect copy number variations (CNVs).

The assay is available in six dyes — FAM, HEX, ATTO 550, ROX, Cy5, and ATTO 700 — and is used at a 20x primer/probe concentration. Wet-lab tested formulations are shipped ready-to-use on dry ice and should be stored protected from light at -30 to -15°C for up to 12 months, or at 2–8°C for up to 6 months, with repeated freeze-thaw cycles avoided. Custom versions are shipped lyophilized at ambient temperature.

The recommended thermal cycling profile is 2 minutes at 95°C for initial heat activation, followed by 40 cycles of 15 seconds at 95°C (denaturation) and 30 seconds at 60°C (combined annealing/extension). For genomic DNA samples with an average length of 20 kb or greater, restriction digestion prior to partitioning is recommended. Template amounts ideally lie within 30–100 ng per reaction and should not exceed 450 ng per reaction when detecting CNV targets present in two copies per diploid genome.