High-Fidelity DNA Polymerase, that provides the highest fidelity amplification currently available, while also offering robust amplification across the broadest range of amplicons, including those that are GC- and AT-rich.
High fidelity amplification is essential for experiments whose outcome depends upon the correct DNA sequence, such as sequencing, SNP detection, protein expression and gene function studies. However, results from such amplifications can be compromised by a DNA polymerase incapable of evenly amplifying diverse types of sequences. For example, high-fidelity PCR of GC-rich regions and other challenging amplicons can lead to lower yields and even non-specific amplification. In addition, any bias in library amplification, associated with next generation sequencing, can lead to challenges in sequence assembly and even "missing" sequence, a problem of growing concern in the sequencing community.
NEB's novel polymerase is fused to the processivity-enhancing Sso7d DNA binding domain, which improves fidelity (more than 50-fold higher than Taq), speed, and performance reliability (amplifying targets up to 20 kb long with extension times as low as 10 seconds per kb). The Q5 buffer system has also been extensively optimized to provide robust performance across both high-GC and high-AT regions, as well as other challenging amplicons.
"High-fidelity polymerases have tended to be more susceptible to difficulties in the PCR amplification of GC-rich or other challenging amplicons. Q5 polymerase has dramatically reduced these limitations and raised the bar for high-fidelity DNA amplification," says Fiona Stewart, Ph.D., Product Marketing Manager at NEB.
A hot start version of the Q5 polymerase, which utilizes a synthetic aptamer for room-temperature set up, is also available. Both the Q5 and Q5 Hot Start DNA Polymerases are available as standalone enzymes or in 2X master mix format for added convenience. A separate formulation has also been developed specifically to meet the needs of amplification of next generation sequencing libraries (NEBNext® High-Fidelity 2X PCR Master Mix).
The discovery and development of high-fidelity polymerases has been, for many years, a key focus of NEB's research. Beginning with the discovery of Vent® DNA Polymerase from Thermococcus litoralis in 1990, and more recently the commercialization of Phusion® DNA Polymerase, NEB researchers have identified and developed a number of high-fidelity polymerases with differing processivities, speeds and degrees of accuracy.
New England Biolabs (NEB) has released a novel DNA polymerase, Q5