HOT FIREPol® Blend Master Mix with 7.5 mM MgCl2

A 5x-concentrated, ready-to-use master mix from Solis BioDyne designed for hot-start PCR with standard cycling. The blend combines HOT FIREPol® DNA polymerase with a proofreading polymerase, providing both 5' flap endonuclease activity and 3'→5' proofreading activity. Compared with HOT FIREPol® used on its own, the blend delivers an increased fidelity of up to five-fold.

Pre-mixed components include the 7.5 mM MgCl2 buffer, dNTPs and BSA; only template, primers and water need to be added by the user. In a 1x reaction the mix supplies 1.5 mM MgCl2 and 200 µM each of dATP, dCTP, dGTP and dTTP. Generated amplicons are compatible with both blunt-end and TA cloning, and treatment with T4 DNA polymerase or the DNA polymerase I large Klenow fragment can be used to raise blunt-end cloning efficiency. The formulation supports amplification of templates up to 5 kb and is recommended for PCR applications that will be visualised by agarose gel electrophoresis with ethidium bromide staining. A suggested thermal profile starts with an initial activation at 95 °C for 12–15 min, followed by 25–30 cycles of denaturation at 95 °C, annealing at 54–66 °C for 30–60 s and extension at 72 °C, with a guideline of about 1 min per kb. Reaction set-up can be performed at room temperature.

Routine storage is at -18 °C to -28 °C; shipping or temporary storage for up to one month at room temperature, or up to six months at 2–8 °C, has no detrimental effect on product quality. Solis BioDyne manufactures the master mix under an ISO 9001 and ISO 13485 certified Quality Management System. The product is supplied for research use only and is not intended for use in diagnostic procedures.