HOT FIREPol® EvaGreen® HRM Mix (no ROX)

Sourced from Solis BioDyne, this 5x concentrated ready-to-use mix is formulated for High Resolution Melt (HRM) Analysis and incorporates EvaGreen® dye. It contains all the components needed to perform qPCR and HRM Analysis — HOT FIREPol® DNA Polymerase, ultrapure dNTPs, MgCl2 and EvaGreen® dye — so the user only adds water, template and primers. The HOT FIREPol® DNA Polymerase is activated by a 12 minute incubation at 95°C, a step that prevents extension of non-specifically annealed primers and primer-dimers formed at low temperatures during qPCR setup.

The mix comprises HOT FIREPol® DNA Polymerase, a 5x EvaGreen® HRM buffer, MgCl2 (12.5 mM in the stock, 2.5 mM in 1x PCR solution), dNTPs, EvaGreen® dye and BSA. Routine storage is at -18°C to -28°C, and shipping or temporary storage for up to 1 month at room temperature has no detrimental effects on the quality of the product. The product is manufactured by Solis BioDyne under ISO 9001 and ISO 13485 certified Quality Management Systems and is supplied for research use only — not for use in diagnostic procedures. The recommended cycling protocol uses an initial activation at 95°C for 12 minutes, then 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.

EvaGreen® is a DNA-binding dye noted for having much less PCR inhibition than SYBR® Green I, for being extremely stable, and for being non-mutagenic and non-cytotoxic. It is compatible with all common real-time PCR cyclers, where standard SYBR® Green or FAM settings can be selected. FIREPol® is an EU registered trademark of Solis BioDyne OÜ, and EvaGreen® is a registered trademark of BIOTIUM, INC.