dPCR Copy Number Assay (200)

QIAGEN offers a digital PCR (dPCR) assay range that enables specific, accurate, reproducible and easy-to-interpret copy number change analysis for an individual gene or region of interest. The assays are supplied in tube format as a ready-to-use 25x-concentrated formulation containing the primer pair for use with the QIAcuity EG PCR Kit and the QIAcuity instrument. More than 200 assays have been dPCR wet-lab tested, with all other in silico designs bench-verified and ready to use in NGS follow-up studies and specific target screening.

Digital PCR is able to detect changes in copy number as small as 1.2 fold or from 5 to 6 copies, enabling accurate identification of aneuploidy in cell lines with chromosomal aberrations previously identified by cytogenetic methods. Detection is based on the intercalating dye EvaGreen. The workflow requires adding genomic DNA, isolated from fresh, frozen or fixed samples, and mastermix from the QIAcuity EG PCR Kit to the individual assays in a QIAcuity Nanoplate. After loading and sealing the nanoplate, run the plate with the recommended cycling program on the QIAcuity instrument; each gene or reference assay is analysed in its own well.

The assays are designed for unique regions of the human genome, and QIAGEN recommends including two reference assays (available separately) to allow accurate calculation of copy number. Copies per microliter are reported through the QIAcuity Software Suite's absolute quantification and copy number data analysis types. Storage is at -20 °C, with a shelf life of 12 months from the manufacturing date. The product is intended for laboratory and research use in copy number variation quantification in DNA samples.