Supreme NZYLong 2× Colourless Master Mix is a premixed readyto-use solution containing Supreme NZYLong DNA polymerase (MB331), an engineered version of NZYLong DNA polymerase designed to amplify longer PCR products, generally of 25 kb and beyond, that displays a higher specificity provided by its hot-startlike PCR capacity. This feature is achieved by a novel hot-start technology, which inhibits both polymerase and 3’→5’ exonuclease activities and thus avoids extension of nonspecifically annealed primers or primer-dimers, as well as degradation of primers and template DNA during PCR reaction setup. The functional activity of the enzyme is restored during a short 5-minute incubation step at 94 °C. The increased processivity of Supreme NZYLong DNA polymerase combined with the hot-start-like PCR capacity results in higher specificity, sensitivity and yield during the amplification of long nucleic acids. The master mix contains dNTPs, reaction buffer and additives at optimal concentrations for the efficient amplification of a wide range of DNA templates. MgCl₂ final concentration is 2.5 mM, allowing the implementation of a variety of PCR protocols. For highly sensitive downstream applications it is recommended to purify the amplified PCR product using NZYGelpure (MB011) before usage in subsequent protocols. Supreme NZYLong DNA polymerase generates a mixture of A-overhang-ended (predominantly) and blunt-ended PCR products, being suitable for cloning with NZYtech’s TA PCR cloning kits (MB053 or MB137).

Extension time: 60 sec/kb at 68 °C

3’→5′ activity: no

Product overhang: mixed