HotStarTaq DNA Polymerase (25000)

This QIAGEN hot-start DNA polymerase is supplied with PCR Buffer and Q-Solution for reaction setup. The enzyme requires an initial heat-activation step of 15 minutes at 95°C before cycling, and remains inactive at room temperature, preventing nonspecific DNA synthesis. The provided 10x PCR Buffer delivers a final concentration of 1.5 mM MgCl2 in the reaction mix, with optional supplementation using 25 mM MgCl2 when higher Mg2+ concentrations are needed.

A typical reaction uses 0.5 µl of the polymerase per 100 µl reaction volume, with recommended cycling of 25–35 cycles including denaturation at 94°C, annealing at 50–68°C, and extension at 72°C. For PCR products longer than 1 kb, an extension time of approximately 1 minute per kb is advised, followed by a final extension of 10 minutes at 72°C. Q-Solution is included to facilitate amplification of templates with high GC content or complex secondary structure, and a No Template Control should always be included.

The polymerase is a modified recombinant form of Thermus aquaticus Taq, cloned in E. coli, suitable for standard PCR, hot-start amplification, SNP locus analysis, methylation-specific PCR, nested PCR, and viral DNA analysis. It exhibits 5'–3' exonuclease activity with an extension rate of 2–4 kb per minute at 72°C and a half-inactivation period of 10 minutes at 97°C or 60 minutes at 94°C. The enzyme should be stored at –30 to –15°C, and post-amplification samples can be held at 2–8°C overnight or at –20°C for longer storage.