New England Biolabs, M0597L, NEBNext® Q5U® Master Mix

NEBNext® Q5U® Master Mix contains a modified version of Q5® High Fidelity DNA Polymerase, optimized for amplification of uracil-containing NGS libraries. This hot start formulation is designed to read through uracils, providing robust, high-fidelity and uniform library amplification. Related Categories Analysis of DNA Methylation by Bisulfite Conversion,, DNA Methylation Analysis,, Epigenetics, Applications Polymerases for NGS Library Preparation,, Specialty PCR FAQ Q: Can NEBNext Q5U Master Mix be used to amplify templates containing uracils? A: Yes. The NEBNext Q5U Master Mix contains Q5U Hot Start High Fidelity DNA Polymerase, which can amplify DNA containing uracils. This master mix can be used with a range of DNA inputs including DNA converted using EM-seq and bisulfite protocols. Q: Is the NEBNext Q5U Master Mix a hot start formulation? A: Yes, the NEBNext Q5U Master Mix is a hot start formulation, and PCR reactions can be set up on ice or at ambient temperature. Q: What cycling conditions should I use for the NEBNext Q5U Master Mix? A: Cycling conditions have been optimized for robust, low-bias amplification of EM-seq libraries. An extension temperature of 65°C is recommended for this application. Q: How many cycles of PCR should I perform with the NEBNext Q5U Master Mix? A: This depends on input amount. When using with the NEBNext Enzymatic Methyl-seq Kit (NEB #E7120), consult the guidelines included in the protocol. Q: Are the DNA products produced by the NEBNext Q5U Master Mix blunt-ended or do they have a single base 3´ overhang? A: As with all Q5 products, NEBNext Q5U Master Mix produces blunt ends. Q: Will NEBNext Ultra II Q5 Master Mix incorporate dUTPs? A: No. NEBNext Ultra II Q5 Master Mix will not incorporate dUTPs and is not compatible with bisulfite-treated DNA. For protocols employing bisulfite-treated DNA, NEBNext(r) Q5U(r) Master Mix (or EpiMark(tm) Hot Start DNA Polymerase) can be used.