NEBNext® Q5 Hot Start HiFi PCR Master Mix is the original hot start NEBNext formulation of Q5 High-Fidelity DNA polymerase. This formulation is also included as a component in all of the original NEBNext Ultra kits, for DNA and RNA library preparation.
However, the NEBNext Ultra II Q5 Master Mix (NEB #M0544) is now available, and this more recent hot start formulation 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® Q5 Hot Start HiFi PCR Master Mix is the original hot start NEBNext formulation of Q5 DNA polymerase. This formulation is also included as a component in all of the original NEBNext Ultra kits, for DNA and RNA library preparation.
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.
This convenient 2X master mix contains dNTPs, Mg++ and a proprietary buffer, and requires only the addition of primers and DNA template for robust amplification. The inclusion of the hot start aptamer allows convenient room temperature reaction set up.
Reaction Conditions NEBNext Q5 Hot Start HiFi
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.
An E. coli strain that carries the Q5 DNA Polymerase gene.
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To ensure optimal performance, themaster mix should be thawed and resuspendedprior to use. Stability testing using up to 30 freeze/thaw cycles has shown no negative effect onmaster mix performance. The NEBNext Q5 HotStart HiFi PCR Master Mix may be liquid at –20°C.
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