HotStarTaq DNA Polymerase (1000 U)

QIAGEN's HotStarTaq DNA Polymerase is a modified form of the recombinant 94 kDa DNA polymerase, originally isolated from Thermus aquaticus and cloned in E. coli, developed to provide hot-start PCR with high specificity. The enzyme is supplied in an inactive state with no polymerase activity at ambient temperatures, which prevents the formation of misprimed products and primer–dimers at low temperatures. Activation requires a 15-minute, 95°C incubation step that can easily be incorporated into existing thermal-cycler programs, and the combination of the enzyme with the unique QIAGEN PCR Buffer minimizes nonspecific amplification products, primer–dimers, and background.

The enzyme is supplied at a concentration of 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 and 60 min at 94°C. It exhibits 5'–3' exonuclease activity and adds an extra A at the 3' end of PCR products, but lacks 3'–5' exonuclease activity. One unit of HotStarTaq DNA Polymerase is defined as the amount of enzyme that will incorporate 10 nmol of dNTPs into acid-insoluble material within 30 min at 72°C. The preparation is free of contaminating nucleases, RNases, proteases, and self-priming activity. The accompanying 10x PCR Buffer contains Tris·Cl, KCl, (NH₄)₂SO₄, and 15 mM MgCl₂ at pH 8.7 (20°C); its balanced combination of KCl and (NH₄)₂SO₄ promotes specific primer–template annealing across a wide range of annealing temperatures and Mg²⁺ concentrations, reducing the need for optimization. A 5x Q-Solution is also included to facilitate amplification of GC-rich templates or templates with extensive secondary structures.

HotStarTaq DNA Polymerase is shipped on dry ice but retains full activity at room temperature (15–25°C) for 2 weeks; for long-term storage it should be kept at –20°C. The product is intended for molecular biology applications and is not intended for the diagnosis, prevention, or treatment of a disease.