HOT FIREPol® Probe Universal qPCR Mix

This 5x master mix from Solis BioDyne is optimized for real-time quantitative PCR and contains all the components necessary to perform singleplex or duplex qPCR, with the exception of template, primers, and probes. A proprietary reaction buffer enables efficient amplification of regular and GC-rich targets, and the formulation is designed for DNA/LNA hydrolysis probes based on the 5' flap endonuclease activity.

An internal passive reference dye based on ROX is included and is compatible with most qPCR cyclers, including platforms that require a high or low ROX reference signal. The mix contains dUTPs to allow UNG treatment for prevention of carryover from previous runs; UNG itself is not included. Components comprise chemically modified FIREPol® DNA Polymerase enabling hot-start with an initial activation of 10 min, a 5x Probe Universal qPCR buffer providing 15 mM MgCl2 (yielding 3 mM MgCl2 in a 1x PCR solution), and dNTPs including dUTP. For multiplex applications, if ROX is used as one of the fluorophores, the internal reference may interfere with the signal.

Suitable applications include DNA/LNA hydrolysis probe-based assays, detection and quantification of DNA and cDNA targets, gene expression profiling, microbial detection, and viral load determination. Routine storage is at -20 °C, and the product can be temporarily stored at room temperature for up to 1 month with no detrimental effect on quality.