HOT FIREPol® Blend Master Mix with 12.5 mM MgCl2

A 5x-concentrated, ready-to-use PCR master mix supplied by Solis BioDyne as a complete formulation containing HOT FIREPol® DNA polymerase, a proofreading enzyme, dNTPs, BSA and an optimized buffer with 12.5 mM MgCl₂ (yielding 2.5 mM at 1x). The enzyme blend provides both 5' flap endonuclease activity and 3'→5' proofreading activity, and the mix exhibits up to five-fold higher fidelity than HOT FIREPol® alone.

The mix is designed for hot-start PCR and is not classified as hazardous under Regulation (EC) No 1272/2008. Generated PCR products are compatible with blunt-end and TA cloning; for improved blunt-end cloning efficiency the products may be treated with T4 DNA polymerase or the DNA polymerase I large Klenow fragment prior to ligation. The formulation is recommended for any PCR application visualised by agarose gel electrophoresis with ethidium bromide staining, and reactions may be assembled at room temperature. Recommended cycling comprises an initial activation at 95°C for 12–15 min, then 25–30 cycles of 95°C for 10–20 s, 54–66°C for 30–60 s and 72°C for 20 s–4 min, followed by a final elongation at 72°C for 5–10 min.

Routine storage is at -28°C to -18°C; shipping and temporary storage for up to 1 month at room temperature or up to 6 months at 2–8°C do not affect product quality. The product is a colorless liquid with a pH of 7.0–9.0 and is manufactured in Tartu, Estonia by Solis BioDyne under an ISO 9001 and ISO 13485 certified Quality Management System. It is supplied for research use only and is covered by patent EP2501716, made by the methods of US Patent No. 9,321,999.