HOT FIREPol® EvaGreen® HRM Mix (no ROX)

A 5x concentrated ready-to-use solution for High Resolution Melt (HRM) analysis, manufactured by Solis BioDyne. The mix incorporates EvaGreen® dye and contains all components required for qPCR and HRM analysis, including HOT FIREPol® DNA Polymerase, ultrapure dNTPs, MgCl2 and EvaGreen® dye — the user only adds water, template and primers. The composition comprises HOT FIREPol® DNA Polymerase, 5x EvaGreen® HRM buffer, 12.5 mM MgCl2 (yielding 2.5 mM MgCl2 in the 1x PCR solution), dNTPs, EvaGreen® dye and BSA.

Reaction setup can be performed at room temperature. The recommended protocol uses 4 µl of the 5x mix in a total reaction volume of 20 µl, with an initial polymerase activation at 95°C for 12 min, followed by 40 cycles of denaturation at 95°C for 15 s, annealing at 60°–65°C for 20 s, and extension at 72°C for 20 s. For templates longer than 150 bp, annealing and extension times may be increased to 30 sec.

EvaGreen® is a DNA-binding dye noted for its low PCR inhibition, high stability, and non-mutagenic, non-cytotoxic profile, and is compatible with all common real-time PCR cyclers using standard SYBR® Green or FAM settings. Routine storage is at -18°C to -28°C; shipping and temporary storage for up to 1 month at room temperature does not affect product quality. The product is manufactured in compliance with ISO 9001 and ISO 13485 certified Quality Management Systems, is intended for in vitro use only, and is for research use only — not for use in diagnostic procedures.