HOT FIREPol® Probe qPCR Mix Plus (ROX)

This 5x probe-based qPCR master mix from Solis BioDyne is optimized for real-time quantitative PCR assays and can amplify up to 2 targets in a single reaction. It contains all components necessary to perform qPCR, with the exception of template, primers, and probe, and includes optimized components together with HOT FIREPol® DNA Polymerase in a proprietary reaction buffer that enables detection of low copy number targets. The mix is designed for DNA hydrolysis probes based on 5' flap endonuclease activity. The polymerase is activated by a 12 min incubation at 95°C, preventing extension of non-specifically annealed primers and primer-dimers formed at low temperatures during qPCR setup.

Typical applications include detection and quantification of DNA and cDNA targets, profiling gene expression, microbial detection, and viral load determination. The mix provides 15 mM MgCl2 (3 mM in a 1x PCR solution) and includes an internal reference based on ROX dye for normalization of the fluorescent reporter signal, with compatibility for both low ROX and high ROX system requirements.

Recommended cycling involves initial activation at 95°C for 12 min, followed by 40 cycles of denaturation at 95°C for 15–20 s and annealing/extension at 60°C for 60 s. The recommended 20 µl reaction uses 4 µl of 5x mix with primers at 200–400 nM and probe at 100–250 nM. Routine storage is at -18°C to -28°C. Manufactured by Solis BioDyne in compliance with the ISO 9001 and ISO 13485 certified Quality Management System, for research use only.