Featured in the ARTIC SARS-CoV-2 sequencing workflow (1) and a component in the NEBNext® ARTIC SARS-CoV-2 kits.
Q5® High-Fidelity DNA Polymerase sets a new standard for both fidelity and robust performance. With the highest fidelity amplification available (~280 times higher than Taq), Q5 DNA Polymerase results in ultra-low error rates. Q5 DNA Polymerase is composed of a novel polymerase that is fused to the processivity-enhancing Sso7d DNA binding domain, improving speed, fidelity and reliability of performance.
Working with uracil-containing DNA templates or using dUTP? Learn about Q5U Hot Start High-Fidelity DNA Polymerase (NEB #M0515).
1. Josh Quick 2020. nCoV-2019 sequencing protocol v3 (LoCost). protocols.io https://www.protocols.io/view/ncov-2019-sequencing-protocol-v3-locost-bh42j8ye
The Q5® Hot Start High-Fidelity 2X
Master Mix features a high-fidelity, thermostable,
hot start DNA polymerase with 3´→ 5´ exonuclease
activity, fused to a processivity-enhancing Sso7d
domain to support robust DNA amplification. The
addition of an aptamer-based inhibitor allows
room temperature reaction setup. With an error rate ~280-fold lower than that of Taq DNA Polymerase, Q5 Hot Start High-Fidelity
2X Master Mix is ideal for cloning and can be used
for long or difficult amplicons. The convenient
master mix formulation is supplied at a 2X
concentration. The mix contains dNTPs, Mg++ and
a proprietary broad-use buffer requiring only the
addition of primers and DNA template for robust
amplification regardless of GC content. When used
at the recommended 1X final concentration, the
Q5 Hot Start High-Fidelity Master Mix contains
2 mM Mg++. Q5 Hot Start High-Fidelity 2X Master
Mix is unlike typical, lower fidelity PCR enzymes. To determine the optimal annealing temperatures for a given set of primers, use of the NEB Tm Calculator is highly recommended.
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