Multiplex PCR NZYProof 2× Green Master Mix is a premixed readyto-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 non-specifically 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. Reactions assembled with Multiplex PCR NZYProof 2× Green Master Mix may be directly loaded onto agarose gels, which simplifies the workflow. There are two dyes (blue and yellow) in the mix that allow monitoring the progress of electrophoresis. Multiplex PCR NZYProof 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 – Multiplex PCR NZYProof 2× Colourless Master Mix (MB338) – or purifying the PCR product using NZYGelpure (MB011) before performing any other protocol.

Extension time: 30 sec/kb at 72 °C

3’→5′ activity: yes

Product overhang: blunt