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

A probe-based one-step RT-qPCR kit from Solis BioDyne, optimized for one-step reverse-transcription real-time quantitative PCR assays. The kit contains all components necessary — except template, primers, and probes — to perform cDNA synthesis and probe-based qPCR in a single tube. Applications include detection and quantification of RNA targets with qPCR and DNA/LNA hydrolysis probe-based assays, and the kit is suitable for singleplex and multiplex reactions. It is compatible with most qPCR cyclers, including those that do or do not require an internal reference dye such as ROX for fluorescent signal normalization.

Reagents include a 5x One-step Multiplex Probe Mix (ROX) with HOT FIREPol® DNA polymerase, reaction buffer, dNTPs (including dUTP), 15 mM MgCl2 (1x qPCR solution — 3 mM MgCl2), and ROX dye; a 40x One-step SOLIScript Mix with SOLIScript Reverse Transcriptase and RiboGrip® RNase Inhibitor; and nuclease-free water. SOLIScript RT is an in silico-engineered thermostable reverse transcriptase active up to 60°C, providing high specificity and working with templates having high secondary structure. HOT FIREPol® DNA Polymerase is activated by a 10-minute incubation at 95°C, preventing extension of non-specifically annealed primers and primer-dimers formed during qPCR setup. If ROX is used as one of the fluorophores, the internal passive reference dye may interfere with the signal — a version without ROX is available.

Storage is at -20°C, with shipping and temporary storage for up to one month at room temperature having no detrimental effect on quality. Recommended cycling conditions: reverse transcription at 50-60°C for 15 min, initial activation at 95°C for 10 min, then 40 cycles of denaturation at 95°C for 15 s and annealing/elongation at 60°C for 60 s. The recommended 20 µl reaction uses 4 µl of 5x probe mix, 0.5 µl of 40x SOLIScript mix, 0.4-0.8 µl of 10 pmol/µl primers (final 200-400 nM), 100-250 nM probe, and up to 20 µl of nuclease-free water, with 1 pg to 1 µg RNA template.