SOLIScript®_ Fast 1-step RT-qPCR Mix with UNG

From Solis BioDyne, this 4x concentrated mix is engineered for sensitive and accurate quantification of RNA targets by one-step real-time RT-PCR using dual-labeled hydrolysis probes (e.g., TaqMan® probes), and is suitable for simultaneous detection and quantitation of up to five targets. The product is supplied in a 1-tube format with nuclease-free water provided in a separate tube and contains all components needed for RT-qPCR apart from the RNA template, primers and probe(s).

The mix includes an in silico engineered thermostable SOLIScript® Reverse Transcriptase active at temperatures up to 60°C, the RNase Inhibitor RiboGrip™ (which inactivates RNase A, RNase B and RNase C), HOT SolisFAST® DNA Polymerase with chemical hot-start and approximately 2–4 times faster extension rates than Taq, and heat-inactivated Salini UNG™ Uracil-N-Glycosylase to prevent carry-over contamination — fully active between 25°C and 40°C and inactive above 50°C. The buffer system provides dNTPs (including dUTPs) with 22.0 mM MgCl2 in the 4x stock (5.5 mM MgCl2 in the 1x RT-qPCR solution).

According to Solis BioDyne's published data, the reagent supports co-detection of RNA, DNA, or both targets in a single reaction and offers inhibitor tolerance and fast cycling compatibility. Demonstrated five-plex RT-qPCR efficiencies on RNA targets were 98.5% (FAM), 96.4% (HEX), 94.4% (ROX), 95.3% (Cy5) and 95.7% (Cy5.5) with R2 ≥ 0.99, while four-plex DNA amplification efficiencies of 100.1% (FAM), 95.5% (HEX), 90.5% (ROX) and 92.3% (Cy5.5) were shown over six 10-fold dilutions of human gDNA from 700 ng/rxn down to 7 pg/rxn. A simultaneous RNA/DNA test system reported efficiencies from 95% to 101% with R2 > 0.99. Recommended input is 1 pg to 1 μg of purified total RNA per 20 µl reaction. Routine storage is at −20°C; the mix is also stable at 4°C for 3 months and at room temperature (25°C) for 1 month.