Do you know what these scientists have in common who say ‘’it is working brilliantly’’, ‘’the reaction worked beautifully’’, ‘’more product’’, ‘’I am surprised by the result’’?
Well, they all use NEB’s Q5 High-Fidelity DNA polymerase for target amplification.
Q5 High-Fidelity DNA Polymerase sets a new standard for both accuracy and robust performance. Regardless of GC content the Q5 DNA Polymerase results in ultra-low error rates along with improved speed, fidelity, and reliability of performance. Through its robust amplification performance amplicons up to 20 kb can be amplified with a high degree of confidence. Moreover, Q5 High-Fidelity DNA Polymerase has one of the highest fidelity amplification rates (>280 times higher than Taq) on the market compared to many other polymerases (Potapov, V. and Ong, J.L. (2017) PLoS ONE. 12(1): e0169774).
Comparison of High-Fidelity Polymerases
For the convenience, Q5 DNA polymerase comes in various formulations such as standalone enzymes, master mix formats or Kits. The Q5 Hot Start DNA polymerase is a thermostable high-fidelity variant of the Q5 DNA polymerase and is an ideal choice for high specificity amplification. It utilizes a unique synthetic aptamer that binds to the polymerase and blocks its activity during the reaction setup. PCR reactions can now be set up at room temperature and the polymerase is activated during normal cycling conditions, thus eliminating the need for a separate high-temperature activation step, which shortens reaction times and improves user friendliness. For instance, Q5 Hot Start High-Fidelity DNA polymerase is part of the LunaScript Multiplex One-Step RT-PCR Kit (NEB #E1555S/L) for cDNA and PCR amplification and enables applications such as diagnostics, pathogen detection, and viral genome sequencing.
The newest formulation of Q5 High-Fidelity DNA Polymerase is the Q5U Hot Start High-Fidelity DNA Polymerase (Q5U). This modified version contains a mutation in the uracil-binding pocket that enables Q5U to read and amplify templates containing uracil and inosine bases. Thus, Q5U is an ideal choice for amplifying bisulfite-converted, enzymatically-deaminated, or damaged DNA. Therefore, it can be concluded Q5 High-Fidelity DNA Polymerase should be the first choice when it comes to fidelity, robustness, coverage, speed, and amplicon length.
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