Not sure why Q5® is your best choice for high-fidelity amplification of GC-rich targets? NEB's scientists will show you why we call Q5 an "ultra-high fidelity polymerase".
Q5 improves on the previous generation of ultra high-fidelity enzymes by offering not only speed, high yields and sensitivity, ultra-low error rates, and the ability to amplify long targets, but also by demonstrating robust performance on very high to very low GC targets.
Q5 is composed of a novel, thermostable DNA polymerase that is fused at the processivity-enhancing Sso7d DNA binding domain.
Q5's unique buffer system and high GC enhancer were designed to provide superior performance with minimal optimization regardless of GC content.
For routine or complex amplicons up to approximately 65% GC content, the Q5 reaction buffer provides reliable and robust amplification.
For amplicons with high GC content, greater than 65% GC, addition of the Q5 high GC enhancer extends the range of this enzyme up to approximately 80% GC content.
For added convenience, Q5 was also developed as a master mix which combines the performance of the standard buffer and high GH enhancer with dNTPs into one easy to use format.
For those preferring room temperature reaction setup, Q5 hot start DNA polymerase takes advantage of a unique aptamer to add hot start convenience without requiring the addition of an activation step.
In addition, an NGS specific formulation of Q5 was developed for low-bias library construction. This formulation has been validated on human and bacterial libraries and provides not only robust library yields and even genome coverage, but also the ultra-high fidelity rates of Q5.
The Q5 product family was developed using a high throughput assay system and a large collection of amplicons from a variety of complex organisms.
Having a quantitative method of evaluating broad PCR performance assisted us not only with the development of robust amplification tools, but also allowed us to critically evaluate other commercial enzymes to ensure that we are providing the best possible reagent available.