SOLIScript® 1-step Multiplex Probe Kit (250 rxn)

A probe-based one-step RT-qPCR reagent kit from Solis BioDyne is optimized for performing cDNA synthesis and probe-based quantitative PCR in a single tube, requiring only template, primers, and probes to be supplied by the user. It is designed for the detection and quantification of RNA targets and is compatible with DNA/LNA hydrolysis probe-based assays in singleplex and multiplex formats.

The kit comprises three tubes: a 5x One-step Multiplex Probe Mix containing HOT FIREPol® DNA polymerase, reaction buffer, dNTPs (including dUTP), 15 mM MgCl₂ (yielding 3 mM at 1x), and an internal reference based on ROX dye; a 40x One-step SOLIScript® Mix containing SOLIScript® Reverse Transcriptase and RiboGrip® RNase Inhibitor; and nuclease-free water. The included reverse transcriptase is engineered for activity at elevated temperatures (up to 60 °C) to support specificity and to handle templates with high secondary structure, while HOT FIREPol® DNA polymerase is activated by a 10 min incubation at 95 °C to limit non-specific amplification and primer-dimer formation.

Compatible with most real-time qPCR cyclers that do or do not require an internal reference dye for normalization, the kit is shipped at room temperature and stored routinely at -20 °C, with temporary storage for up to one month at room temperature reported to have no detrimental effect on quality. A recommended cycling protocol in a 20 µl reaction includes reverse transcription at 50–60 °C for 15 min, initial activation at 95 °C for 10 min, and 40 cycles of denaturation at 95 °C for 15 s followed by annealing/elongation at 60 °C for 60 s.