HOT FIREPol® Probe Universal qPCR Mix

HOT FIREPol^® Probe Universal qPCR Mix is optimized for real-time quantitative PCR assays and contains all the components necessary to perform singleplex or duplex qPCR, with the exception of template, primers, and probes. Proprietary reaction buffer that enables efficient amplification of regular and GC-rich targets.

HOT FIREPol^® Probe Universal qPCR Mix is optimized for DNA/LNA hydrolysis probes based on the 5′ flap endonuclease activity.

The Mix contains internal passive reference dye that is compatible with most qPCR cyclers including high ROX or low ROX reference signal requiring platforms.

The Mix contains dUTPs to prevent cross-contamination with UNG treatment.

Applications

• DNA/LNA hydrolysis probe-based assays
• Detection and quantification of DNA and cDNA targets
• Profiling gene expression
• Microbial detection
• Viral load determination

Properties

Concentration: 5x

Hot-start: yes, initial activation in 10 min

Detection type: Probe-based

Reference dye: reference dye based on ROX

Compatible real-time instruments: Most qPCR platforms. Check Your cycler!

Mix Components

HOT FIREPol^® DNA Polymerase: chemically modified FIREPol^® DNA Polymerase enabling hot-start

5x Probe Universal qPCR buffer with 15 mM MgCl₂: 1x PCR solution – 3 mM MgCl₂

dNTPs including dUTP:  Mix allows UNG treatment to prevent carryover contamination from previous runs.
IMPORTANT: UNG is not included in the HOT FIREPol^® Probe Universal qPCR Mix

Reference dye based on ROX: For multiplex application – if ROX dye is used as one of the fluorophores, the internal reference might interfere with the signal.

In separate vial

100% DMSO is included in the kit in a separate vial. DMSO is recommended as a PCR additive for templates with high GC content. In some cases, DMSO is also required to relax secondary structures.