HotStarTaq DNA Polymerase (5000)

From QIAGEN, this hot-start DNA polymerase is a modified form of a recombinant 94 kDa DNA polymerase originally isolated from Thermus aquaticus and cloned in E. coli (Deoxynucleoside-triphosphate: DNA deoxynucleotidyltransferase, EC 2.7.7.7). It 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, 95°C incubation step that can be incorporated into existing thermal cycling programs. Reaction setup is quick and convenient because all reaction components can be combined at room temperature.

The enzyme is supplied at a concentration of 5 units/µl. One unit is defined as the amount of enzyme that will incorporate 10 nmol of dNTPs into acid-insoluble material within 30 minutes at 72°C. The extension rate is 2–4 kb/min at 72°C, and the half-life is 10 min at 97°C and 60 min at 94°C. The polymerase exhibits 5'–3' exonuclease activity and adds an extra adenosine (A) to amplification products, but lacks 3'–5' exonuclease activity as well as nuclease, protease, RNase, and self-priming activity. Substrate analogs accepted include dNTP, ddNTP, and fluorescent dNTP/ddNTP.

Designed to provide higher PCR specificity, this polymerase minimises nonspecific amplification products, primer–dimers, and background, and is well suited to amplification of complex genomic or cDNA templates, very low-copy targets, or assays using multiple primer pairs. The accompanying 10x PCR Buffer contains Tris·Cl, KCl, (NH4)2SO4, and 15 mM MgCl2 at pH 8.7 (20°C), with a 5x Q-Solution and a 25 mM MgCl2 solution also provided. The product is shipped on dry ice but retains full activity at room temperature (15–25°C) for 2 weeks, and should be stored immediately at –20°C in a constant temperature freezer upon receipt. It is developed, designed, and sold for research purposes only and is not to be used for human diagnostic or drug purposes unless expressly cleared for that purpose by the appropriate regulatory authorities.