QIAcuity OneStepAdvanced Probe Kit (1mL)

QIAGEN provides a one-step RT-PCR kit for the QIAcuity digital PCR instruments, enabling sensitive quantification of RNA or RNA+DNA targets in a single combined reverse transcription and PCR reaction. The kit contains a 4x concentrated Master Mix and a 100x concentrated RT Mix optimized for microfluidic use with QIAcuity Nanoplates, enhancing the specificity and efficiency of probe-based digital PCR to deliver accurate singleplex or up to 5-plex analysis. The included HotStart Reverse Transcription (RT) Enzyme remains non-functional at room temperature, preventing generation of off-target cDNA even during extended incubation at 37°C, allowing reaction setup at ambient temperature and parallel loading of multiple Nanoplates into QIAcuity Four and Eight instruments.

An optional Enhancer GC supplied in a separate vial is recommended when amplicons are GC-rich or longer than 150 nt, and for use with TaqMan (Thermo Fisher) gene expression assays. The kit also includes a QuantiNova Internal Control RNA (QN IC RNA) that can be used optionally as a reverse transcription and amplification control. It uses QIAGEN's QuantiNova DNA Polymerase paired with proprietary buffer technology optimized for Nanoplate microfluidics, delivering consistent performance in terms of sensitivity, reproducibility, and efficiency. The ready-to-use master mix eliminates the need to optimize reaction and cycling conditions; users add the 100x RT mix, template RNA, and primer-probe sets and follow the Quick-Start Protocol. Combined with the QIAcuity Digital PCR System and QIAcuity Nanoplates, the kit supports applications such as viral RNA and mRNA detection, as well as simultaneous RNA and DNA target analysis — for example, wastewater testing where a mixture of four bacterial gDNAs (Clostridioides difficile, Vibrio cholerae, Yersinia enterocolitica, Listeria monocytogenes) and SARS-CoV-2 RNA has been used as input with 26k 24-well Nanoplates at template dilutions of 500, 50 and 5 copies/μl. Manufacturer-reported ABI TaqMan Assays values include ATOX1 at 100%, 68%, and 95% and TGFBR2 at 100%, 87%, and 107%.