HOT FIREPol® Multiplex qPCR Mix (ROX)

A 5x multiplex qPCR master mix from Solis BioDyne, optimized for probe-based real-time quantitative PCR and capable of amplifying up to 4 targets in a single reaction. The mix contains HOT FIREPol® DNA Polymerase, optimized buffer components, ultrapure dNTPs (including dUTP), MgCl2 and ROX passive reference dye, providing all components except primers, probes and template. It is specifically designed for DNA hydrolysis probes based on the 5' flap endonuclease activity.

Applications include detection and quantification of DNA and cDNA targets, gene expression profiling, microbial detection, and viral load determination. The 5x buffer contains 15 mM MgCl2, yielding a final concentration of 3 mM MgCl2 in a 1x reaction. The mix includes dUTP, allowing optional UNG treatment to prevent carryover contamination, though UNG must be purchased separately. The ROX passive reference dye is compatible with both low ROX and high ROX system requirements.

The recommended qPCR cycling protocol begins with an initial activation at 95°C for 10 minutes, followed by 40 cycles of denaturation at 95°C for 15–20 seconds and annealing/extension at 60°C for 60 seconds. Forward and reverse primers are recommended at 200–400 nM each, with probes at 100–250 nM. Suggested template concentrations are 0.1 pg/µl–10 ng/µl for cDNA and 10 pg/µl–4 ng/µl for gDNA. The product is manufactured under ISO 9001 and ISO 13485 certified Quality Management Systems, is for research use only, and should be stored at -18°C to -28°C. Shipment and temporary storage for up to 1 month at room temperature have no detrimental effects on quality.