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

A 5x-concentrated ready-to-use PCR master mix supplied by Solis BioDyne as a green liquid that contains all reagents required for PCR except template, primers and water. The blend combines HOT FIREPol® DNA polymerase with a proofreading polymerase, providing both 5' flap endonuclease activity and 3'→5' proofreading activity, with up to five-fold higher fidelity than HOT FIREPol® alone. It includes 10 mM MgCl2 (yielding 2 mM MgCl2 in 1x solution), 1 mM dNTPs (200 µM of each in 1x), BSA, two tracking dyes — blue (migration equivalent to 3.5–4.5 kb DNA fragment) and yellow (migration equivalent to 35–45 bp DNA fragment) — and a compound that increases sample density so reactions can be loaded directly onto agarose gels. The liquid has a pH of 7.0–9.0.

Generated PCR products are compatible with blunt-end and TA cloning procedures. The mix is recommended for hot-start PCR applications visualised by agarose gel electrophoresis with ethidium bromide staining, but is not recommended for spectrophotometric (absorbance or fluorescence) measurements, since the dyes can interfere. Routine storage is at -28ºC to -18ºC; shipping or temporary storage for up to 1 month at room temperature, or up to 6 months at 2–8ºC, has no detrimental effect on quality. A typical PCR cycling protocol starts with initial activation at 95ºC for 12–15 minutes, followed by 25–30 cycles of denaturation at 95ºC for 15–20 s, annealing at 54–66ºC for 30–60 s and extension at 72ºC for 20 s–4 min, with a final extension at 72ºC for 5–10 min. The product is for research use only and is manufactured by Solis BioDyne in compliance with ISO 9001 and ISO 13485 certified Quality Management System.