HOT FIREPol® GC Master Mix

From Solis BioDyne, this 5x-concentrated ready-to-use master mix has been developed for PCR amplification of difficult GC-rich templates and DNA secondary structures, with demonstrated performance on templates up to 79% GC content. The kit includes 100% DMSO and 25 mM MgCl2 supplied in separate vials for added flexibility, and its special buffer is optimized for high-yield GC-rich amplification.

The mix contains a chemically modified FIREPol® DNA Polymerase that enables hot-start activation, together with the 5x HOT FIREPol® GC Buffer (providing 7.5 mM MgCl2, equivalent to 1.5 mM in a 1x PCR solution), dNTPs, and BSA. All reagents required for PCR are provided except template DNA, gene-specific primers, and nuclease-free PCR-grade water. A typical 20 µl reaction uses 4 µl of the 5x master mix; DMSO is recommended as a PCR additive for templates with high GC content and can be optimized in 2.5% increments up to 10%.

Polymerase activation requires an initial 95°C incubation for 12 minutes, followed by 25–30 cycles of denaturation at 95°C (15–30 sec), annealing at 54–66°C (30–60 sec), and extension at 72°C (1.5–5.5 min), with a final extension at 72°C for 5–10 min. The reagents are shipped at room temperature and the reaction can be set up at room temperature. Routine storage is at –20°C until the expiry date; the product is stable at 4°C for 6 months and at room temperature (25°C) for 1 month, with freeze-thaw stability of 30 cycles. 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.