Multiplex PCR NZYProof 2× Colourless Master Mix is a premixed ready-to-use solution designed for the simultaneous amplification of multiple DNA fragments (up to 15 targets) in a single tube with high accuracy. The Master Mix contains an engineered proofreading DNA polymerase displaying a hot-start-like PCR capacity and high processivity (fast polymerisation), while reducing errors due to its 3´→5´ exonuclease activity. The enzyme is inactive at room temperature, avoiding extension of nonspecifically annealed primers or primer-dimers, and thus ensuring highly specific and sensitive multiplex PCR amplification. In addition, the master mix contains dNTPs and reaction buffer at optimal concentrations for efficient multiplexing of targets ranging in size from 70 bp to 2.5 kb over a broad range of primer and template concentrations. MgCl2 final concentration is 1.5 mM, allowing the implementation of a comprehensive variety of PCR protocols.

Extension time: 30 sec/kb at 72 °C

3’→5′ activity: yes

Product overhang: blunt