Supreme NZYTaq II 2× Green Master Mix is a premixed ready-to-use solution containing Supreme NZYTaq II DNA polymerase (MB355), a robust DNA polymerase derived from Taq DNA polymerase that was engineered to exhibit high processivity and to provide high PCR sensitivity. The enzyme, displaying a hot-start-like PCR capacity, is inactive at room temperature, avoiding extension of non-specifically annealed primers or primer-dimers and providing a highly specific DNA amplification. The functional activity of the enzyme is restored during a short 5-minute incubation step at 95 °C. In addition, the master mix contains dNTPs, reaction buffer and additives at optimal concentrations for the efficient amplification of a wide range of DNA templates of up to 6 kb. MgCl2 final concentration is 2.5 mM, allowing the implementation of a variety of PCR protocols. In addition, reactions assembled with Supreme NZYTaq II 2× Green Master Mix may be directly loaded onto agarose gels. The mix contains two dyes (blue and yellow) that allow monitoring the progress of the electrophoresis. Supreme NZYTaq II 2× Green Master Mix is not suitable when direct fluorescent or absorbance readings are required without prior purification of the amplified DNA from PCR. We recommend using the master mix version without dyes Supreme NZYTaq II 2× Colourless Master Mix (MB359) or purifying the PCR product using NZYGelpure (MB011) before performing such downstream protocols. Supreme NZYTaq II DNA polymerase lacks 3’→5′ exonuclease activity. Resulting PCR products have an A-overhang and are suitable for cloning with NZYtech’s NZY-A PCR cloning kit (MB053) or NZY-A Speedy PCR cloning kit (MB137).

Extension time: 15-30 sec/kb at 72 °C

3’→5′ activity: no

Product overhang: 3′-A