NEBNext® High-Fidelity 2X PCR Master Mix
However, the NEBNext Ultra II Q5 Master Mix (NEB #M0544) is now available, and this more recent formulation is a hot start and also further improves the uniformity of amplification of libraries, including superior performance with GC-rich regions. The NEBNext Ultra II Q5 Master Mix is also included as a component in all NEBNext Ultra II kits, for DNA and RNA library preparation.
NEBNext High-Fidelity 2X PCR Master Mix is the original non-hot start NEBNext formulation of Q5 DNA Polymerase. This formulation is also included as a component in the original NEBNext standard workflow library prep kits for Illumina, and in NEBNext library prep kits for Ion Torrent.
The NEBNext Ultra II Q5 Master Mix (NEB #M0544) is now available, which further improves the uniformity of amplification of libraries, including superior performance with GC-rich regions.
The polymerase component of both master mixes, Q5 High-Fidelity DNA Polymerase, is a novel thermostable DNA polymerase that possesses 3´→5´ exonuclease activity, and is fused to a processivity-enhancing Sso7d domain. Q5 also has the highest fidelity available (> 280-fold higher than that of Taq DNA Polymerase), resulting in ultra-low error rates.
NEBNext High-Fidelity 2X PCR Master Mix contains dNTPs, Mg++ and a proprietary buffer, and requires only the addition of primers and DNA template for robust amplification.
For larger volume requirements, customized and bulk packaging is available by purchasing through the Custom Solutions department at NEB. Please contact email@example.com for further information.
Product SourceAn E. coli strain that carries the Q5 High-Fidelity DNA Polymerase gene.
Reaction Volume DefinitionNEBNext High-Fidelity 2X PCR Master Mix, DNA template and 0.5 μM to 1.25 μM primers (depending on sample input) in a total reaction volume of 50 μl.
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