HotStarTaq Master Mix Kit (1000 U)

QIAGEN supplies this ready-to-use 2x master mix formulated for hot-start PCR, combining HotStarTaq DNA Polymerase, PCR Buffer, and dNTPs in a single tube to minimize hands-on time and variability between reactions. HotStarTaq DNA Polymerase is provided in an inactive state at ambient temperatures, preventing the formation of misprimed products and primer–dimers, and is activated by a 15-minute incubation at 95°C that can be incorporated directly into standard thermal cycling programs. Each 50 µl reaction set up from the 2x master mix delivers 2.5 units of HotStarTaq DNA Polymerase, 200 µM of each dNTP, and a final MgCl₂ concentration of 1.5 mM.

The enzyme is a modified form of a recombinant 94 kDa DNA polymerase originally isolated from Thermus aquaticus and cloned in E. coli, supplied at 5 units/µl with an extension rate of 2–4 kb/min at 72°C and a half-life of 10 min at 97°C or 60 min at 94°C. It possesses 5′–3′ exonuclease activity and adds an extra A to amplified products, while showing no detectable 3′–5′ exonuclease, nuclease, protease, RNase, or self-priming activity. The master mix is shipped on dry ice, retains full activity at room temperature (15–25°C) for 3 days, and should be stored upon receipt at –30 to –15°C in a constant-temperature freezer.

Designed for research use only, the kit is well suited to amplifying complex genomic or cDNA templates, very low-copy targets, and reactions using multiple primer pairs. Because the polymerase remains inactive at room temperature, reaction setup does not require keeping PCR tubes on ice, although a No Template Control should always be included.