HOT FIREPol® Blend Master Mix Ready to Load with 10 mM MgCl2

A 5x-concentrated ready-to-load master mix from Solis BioDyne provides all reagents required for PCR except template, primers, and water. The formulation includes a density compound that allows direct loading onto agarose gels, plus two tracking dyes — a blue dye that migrates equivalent to a 3.5–4.5 kb DNA fragment and a yellow dye that migrates equivalent to a 35–45 bp DNA fragment — so progress can be monitored during electrophoresis.

The enzyme blend pairs HOT FIREPol® DNA polymerase with a proofreading polymerase and delivers up to five-fold higher fidelity compared with HOT FIREPol® used alone. The 5x buffer contains 10 mM MgCl2, giving a final 2 mM MgCl2 in the 1x PCR solution, and 200 µM each of dATP, dCTP, dGTP, and dTTP in the 1x working concentration, together with BSA. The mix is intended for hot-start PCR applications where amplicons will be visualised by agarose gel electrophoresis with ethidium bromide staining, and is not recommended for spectrophotometric workflows because the dyes can interfere with absorbance or fluorescence measurements.

Reaction setup at room temperature is recommended, and an initial 95°C step for 12–15 minutes is required to activate the polymerase. A representative 20 µl reaction uses 4 µl of the 5x master mix with 0.1–0.3 µM each primer and 5–50 ng/µl template DNA. Solis BioDyne manufactures the product under ISO 9001 and ISO 13485 certified quality management systems, for research use only.