HOT FIREPol® EvaGreen® qPCR Mix Plus (no ROX)

A 5x ready-to-use master mix from Solis BioDyne, the HOT FIREPol® EvaGreen® qPCR Mix Plus (no ROX) is optimised for real-time quantitative PCR (qPCR) assays using dye-based detection. The mix incorporates EvaGreen® dye and contains all components necessary to perform qPCR, including HOT FIREPol® DNA Polymerase, ultrapure dNTPs, MgCl2, and EvaGreen® dye, so the user only needs to add water, template, and primers. HOT FIREPol® DNA Polymerase is activated by a 12 min incubation step at 95°C, which prevents extension of non-specifically annealed primers and primer-dimers formed at low temperatures during reaction setup.

The mix is suited to detection and quantification of DNA and cDNA targets, profiling gene expression, microbial detection, and viral load determination. The composition includes HOT FIREPol® DNA Polymerase, 5x EvaGreen® qPCR buffer (12.5 mM MgCl2, giving 2.5 mM in a 1x PCR solution), dNTPs, and EvaGreen® dye. The recommended qPCR cycling protocol starts with an initial activation at 95°C for 12 minutes, followed by 40 cycles of denaturation at 95°C for 15 seconds, annealing at 60°–65°C for 20 seconds, and extension at 72°C for 20 seconds.

EvaGreen® is a DNA-binding dye with spectra similar to SYBR® Green I, but has much less PCR inhibition, is an extremely stable dye, and has been shown to be non-mutagenic and non-cytotoxic. It is compatible with all common real-time PCR cyclers when standard SYBR® Green or FAM settings are selected. Routine storage is at -18°C to -28°C, while shipping or temporary storage at room temperature for up to 1 month has no detrimental effects on product quality. The product is manufactured by Solis BioDyne in compliance with ISO 9001 and ISO 13485 certified Quality Management Systems and is supplied for research use only.