HOT FIREPol® Blend Master Mix with 7.5 mM MgCl2

A 5x-concentrated, ready-to-use hot-start PCR master mix from Solis BioDyne, the formulation combines HOT FIREPol® DNA polymerase with a proofreading enzyme to deliver both 5' flap endonuclease and 3'→5' proofreading activities. Compared with HOT FIREPol® alone, the blend shows up to five-fold higher fidelity and is suitable for templates up to 5 kb. PCR products are compatible with blunt-end and TA cloning; treating the products with T4 DNA polymerase or DNA polymerase I large Klenow fragment can further improve blunt-end cloning efficiency.

The mix is supplied at 7.5 mM MgCl2 (giving 1.5 mM MgCl2 in a 1x reaction) and contains 1 mM of each dNTP (200 µM each in 1x), BSA, and 5x Blend Master Mix Buffer. The manufacturer recommends it for any PCR application to be visualized by agarose gel electrophoresis and ethidium bromide staining, and highlights benefits including increased yield, sensitivity and specificity, up to 5x higher fidelity, reduced primer dimer formation, and reaction set-up and shipment without ice, supported by the proprietary Stability TAG technology. A Ready To Load version is also offered.

Routine storage is at -18°C to -28°C; temporary storage of up to 1 month at room temperature, or up to 6 months at 2–8°C, has no detrimental effects on quality. The product is manufactured under an ISO 9001 and ISO 13485 certified Quality Management System and is for research use only. A recommended PCR protocol uses an initial activation of 95°C for 12–15 min, followed by 25–30 cycles of denaturation at 95°C for 10–20 s, annealing at 54–66°C for 30–60 s, and elongation at 72°C for 20 s to 4 min, with a final elongation at 72°C for 5–10 min.