New England Biolabs, M0541S, NEBNext® High-Fidelity 2X PCR Master Mix

Applications Polymerases for NGS Library Preparation,, Fast PCR,, High-Fidelity PCR, Specification Heat Inactivation No FAQ Q: What are the advantages of using NEBNext High-Fidelity 2X PCR Master Mix? A: NEBNext High-Fidelity 2X PCR Master Mix provides a unique combination of ultra-high fidelity and robust amplification of NGS libraries, regardless of GC content. Q: Is NEBNext® High-Fidelity 2X PCR Master Mix the same as the Q5® High-Fidelity Master Mix? A: No. Although both master mixes contain Q5 High-Fidelity DNA Polymerase, the master mix composition is different. NEBNext High-Fidelity 2X PCR Master Mix has been specifically optimized for amplification of NGS libraries, while Q5 High-Fidelity Master Mix has been optimized for amplification of a variety of templates. Q: Do I need to make any changes to the cycling conditions for my NGS library amplification I have previously been using with Phusion High-Fidelity PCR Master Mix? A: No. cycling conditions used successfully with Phusion High-Fidelity PCR Master Mix can also be used for NEBNext High-Fidelity 2X PCR Master Mix. Q: How many cycles of PCR should I perform with NEBNext High-Fidelity 2X PCR Master Mix? A: This depends on input amount. For use with NEBNext kits, consult the guidelines included in the product manuals. Q: Will NEBNext High-Fidelity 2X PCR Master Mix incorporate dUTPs? A: No. NEBNext High-Fidelity 2X PCR Master Mix will not incorporate dUTPs and is not compatible with bisulfite-treated DNA. For protocols employing bisulfite-treated DNA EpiMark™ Hot Start DNA Polymerase can be used. Q: Are the DNA products produced NEBNext High-Fidelity 2X PCR Master Mix blunt-ended or do they have a single base 3’ overhang? A: NEBNext High-Fidelity 2X PCR Master Mix produces blunt ends.