HOT FIREPol® Multiplex qPCR Mix (ROX)

From Solis BioDyne, this 5x probe-based master mix is designed for multiplex quantitative PCR, allowing simultaneous amplification and quantification of up to four DNA or cDNA targets in a single reaction. Combining several targets in one well reduces reagent cost and reaction set-up time, and the formulation is reported as robust with GC-rich templates. The mix is supplied as a 5x concentrate containing all components required for qPCR except primers, probes and template, and includes an internal ROX passive reference dye for signal normalization on both low-ROX and high-ROX instruments.

The reagent combines HOT FIREPol® DNA Polymerase with a 5x Multiplex qPCR buffer, ultrapure dNTPs (including dUTP) and MgCl2 at 15 mM in the 5x stock (3 mM in a 1x reaction). The hot-start mechanism is activated by a 10-minute incubation at 95 °C, which limits non-specific primer extension and primer-dimer formation during reaction set-up, and the mix is optimized for DNA hydrolysis probes using 5' flap endonuclease activity. Because dUTP is included, the mix supports UNG carryover-prevention treatment; UNG itself is not included and must be purchased separately. A version with Purple reference dye is also available for users who want to reserve the ROX channel for target detection.

Typical applications include detection and quantification of DNA and cDNA targets, gene expression profiling, microbial detection, and viral load determination. Recommended template input ranges are cDNA at 0.1 pg/µl–10 ng/µl and gDNA at 10 pg/µl–4 ng/µl. The suggested cycling protocol begins with a 10-minute 95 °C activation, followed by 40 cycles of 95 °C denaturation for 15–20 s and 60 °C annealing/extension for 60 s. The product is shipped without ice and remains stable at room temperature for up to 30 days, with routine storage at -18 °C to -28 °C. It is manufactured under ISO 9001 and ISO 13485 quality management systems and is intended for research use only.