FIREPol® Master Mix (12.5 mM MgCl2)

A ready-to-use 5x master mix from Solis BioDyne containing FIREPol® DNA polymerase and all reagents required for PCR except template, primers and water. The mix is supplied with 12.5 mM MgCl2, giving a final concentration of 2.5 mM MgCl2 in a 1x PCR solution. Each dNTP is present at 1 mM, providing 200 µM of dATP, dCTP, dGTP and dTTP in the 1x reaction. The 5x Reaction Buffer B contains 0.4 M Tris-HCl, 0.1 M (NH4)2SO4 and 0.1% w/v Tween-20. It is suited for a wide range of PCR assays and TA cloning.

For a 20 µl reaction, 4 µl of the 5x master mix is combined with 0.2-0.6 µl each of forward and reverse primers at 10 pmol/µl for a final concentration of 0.1-0.3 µM, 5-50 ng/µl of template DNA, and water to volume. Recommended cycling conditions are: initial denaturation at 95°C for 3-5 min (1 cycle); denaturation at 95°C for 20-40 s; annealing at 54-66°C for 30-60 s; elongation at 72°C for 40 s to 4 min (25-30 cycles); and final elongation at 72°C for 5-10 min. The mix is recommended for PCR applications visualized by agarose gel electrophoresis and ethidium bromide staining, and is not recommended where a detergent-free buffer system is required. Routine storage is at -20°C; shipping and temporary storage for up to 1 month at room temperature or up to 6 months at 2-8°C has no detrimental effect on quality.

For in vitro use only. The product and its components should be handled only by persons trained in laboratory techniques. Suitable protective clothing such as laboratory overalls, gloves and safety glasses is advisable. In case of contact with skin or eyes, wash immediately with water. Some applications may require a license which is not provided by the purchase of this product; users should obtain the license if required.