HOT FIREPol® EvaGreen® HRM Mix (no ROX)

A 5x concentrated, ready-to-use solution from Solis BioDyne designed for High Resolution Melt (HRM) analysis. The mix is supplied without ROX dye and contains all components required to perform qPCR and HRM, including HOT FIREPol® DNA Polymerase, ultrapure dNTPs, MgCl2 and EvaGreen® dye. Users only need to add PCR-grade water, template and primers.

Composition includes HOT FIREPol® DNA Polymerase, 5x EvaGreen® HRM buffer containing 12.5 mM MgCl2 (yielding 2.5 mM MgCl2 in the 1x PCR solution), dNTPs, EvaGreen® dye and BSA. The included HOT FIREPol® DNA Polymerase is activated by a 12 minute incubation at 95°C, which prevents extension of non-specifically annealed primers and primer-dimers formed at low temperatures during qPCR setup. Recommended cycling is initial 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. Reaction setup at room temperature is recommended.

The EvaGreen® dye is described as a DNA-binding dye with much less PCR inhibition than SYBR® Green I, extreme stability and a non-mutagenic, non-cytotoxic profile. It is compatible with all common real-time PCR cyclers using standard SYBR® Green or FAM settings. The product should be routinely stored between -18°C and -28°C, and tolerates up to 1 month at room temperature during shipping without detrimental effects on quality. It is manufactured under ISO 9001 and ISO 13485 certified Quality Management Systems and is for research use only. FIREPol® is an EU registered trademark of Solis BioDyne OÜ; EvaGreen® is a registered trademark of Biotium, Inc.