QIAcuity HEK293 resDNA Quant Kit (96)

From QIAGEN, this residual DNA quantification kit uses the QIAcuity digital PCR system to detect trace levels of HEK293 host cell DNA in biopharmaceutical products. It is designed for bioprocess manufacturing and quality control, addressing FDA and WHO guidelines on residual DNA limits. The assay targets a multicopy, undefined repeated-element locus with an amplicon size below 100 bp, making quantification unaffected by fragmentation level. The kit ships on dry ice and is stored at −30 to −15°C. Components include HEK293 resDNA Quant Master Mix (2x) supplied in lyophilized format, an internal control assay detected in the Green and Yellow channels, internal control DNA, positive control DNA, and dPCR qualified water. The recommended reaction setup uses 20 µL Master Mix, 1 µL internal control (100 ± 20 copies/µL final concentration), restriction enzyme at 0.025−0.25 U/µL, and dPCR qualified water to a total reaction volume of 40 µL. With Nanoplate 26k (24-well), up to 28 µL of template can be loaded per reaction. DNA samples with average length ≥20 kb should be restriction-digested before partitioning. Thermal cycling is 2 min at 95°C initial activation, 5 two-step cycles (15 s at 95°C, 30 s at 60°C), and 40 two-step cycles (15 s at 95°C, 30 s at 66°C). Imaging uses the Green channel for the HEK293 target and the Yellow channel for the internal control, both with 500/6 exposure/gain. The kit shows high linearity from 0.5–8,000 pg per reaction. A copies/µL to fg/µL conversion factor of 1.54 is provided. DNA extraction is not required, but is recommended for samples with high inhibitor or protein levels.