The NEBNext Q5 Hot Start HiFi PCR Master Mix is specifically optimized for robust, high-fidelity amplification of next-generation sequencing (NGS) libraries, regardless of GC content.
The polymerase component of the master mix, 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 and ultra-low error rate (>100-fold lower than that of Taq DNA Polymerase and ~12-fold lower than that of Pyrococcus furiosus (Pfu) DNA Polymerase. The buffer component of the master mix has been optimized for robust amplification, even with GC-rich regions, and offers enhanced compatibility with a variety of beads used in typical NGS workflows, as well as improved tolerance to over-cycling. These features make the NEBNext Q5 Hot Start HiFi PCR Master Mix ideal for NGS library amplification.
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.