HOT FIREPol® Probe qPCR Mix Plus (ROX)

Solis BioDyne supplies a 5x probe-based qPCR master mix optimized for real-time quantitative PCR using sequence-specific fluorogenic DNA-probe chemistries. The mix is designed for amplifying up to two targets in a single reaction and contains all components necessary to perform qPCR with the exception of template, primers, and probe. It is compatible with both low-ROX and high-ROX system requirements, relying on an internal reference based on ROX dye to normalize the fluorescent reporter signal.

The mix includes HOT FIREPol^® DNA Polymerase supplied in a proprietary 5x Probe qPCR buffer with 15 mM MgCl₂ (3 mM MgCl₂ in a 1x PCR solution) and dNTPs. The polymerase is activated by a 12 min incubation at 95 °C, which prevents extension of non-specifically annealed primers and primer-dimers formed at low temperatures during qPCR setup. The mix is suited to DNA hydrolysis probes based on 5' flap endonuclease activity, with applications including detection and quantification of DNA and cDNA targets, gene expression profiling, microbial detection, and viral load determination.

Recommended reaction setup is performed at room temperature, with a 20 µl total volume containing 4 µl of the 5x mix, 200–400 nM each primer, and 100–250 nM probe. The recommended cycling protocol is 95 °C for 12 min, followed by 40 cycles of 95 °C for 15–20 s and 60 °C for 60 s. Routine storage is at -18 °C to -28 °C; shipping and temporary storage for up to one month at room temperature has no detrimental effect on quality. The product is manufactured by Solis BioDyne in compliance with ISO 9001 and ISO 13485 and is for research use only.