5x HOT FIREPol® Probe Universal qPCR Mix, 250 reactions

5x HOT FIREPol^® Probe Universal qPCR Mix is optimized for real-time quantitative PCR assays and contains all the components necessary to perform qPCR, with the exception of template, primers, and probe. The qPCR Mix contains optimized components and HOT FIREPol® DNA Polymerase supplied in a proprietary reaction buffer that enables efficient amplification of AT-rich, regular and GC-rich (up to 75%) targets. HOT FIREPol^® Probe Universal qPCR Mix is optimized for DNA/LNA hydrolysis probes based on the 5′ flap endonuclease activity.

HOT FIREPol^® DNA Polymerase is activated by a 10 min incubation step at 95°C. This prevents extension of non-specifically annealed primers and primer-dimers formed at low temperatures during qPCR setup.

Features & Benefits

• increased sensitivity and specificity for wide range of templates, including AT-rich, regular and GC-rich cDNA and gDNA
• suitable for singleplex and duplex assays
• reaction set-up at room temperature -master mix is stable at ambient temperature for one month
• mix contains dUTP – it may be used with UNG to prevent carryover contamination from previous qPCR runs (UNG treatment)
• use with any hydrolysis probe (LNA or DNA)
• wide instrument compatibility: the mix is compatible with any real-time instrument other then capillary

Applications

• DNA/LNA hydrolysis probe-based assays
• detection and quantification of DNA and cDNA targets
• profiling gene expression
• microbial detection
• viral load determination

Mix Components

• HOT FIREPol^® DNA Polymerase
• 5x Probe Universal qPCR buffer
• 15 mM MgCl₂ 1x PCR solution – 3 mM
• dNTPs blend containing dUTP/dTTP
• Internal reference based on ROX dye

Shipping & Storage

Routine storage: -20°C

Temporary storage for up to 1 month at room temperature has no detrimental effects on the quality of the product.

Full Protocol