FAQ: What buffers are recommended for shearing DNA in NEB’s enzymatic methyl-seq (EM-seq) workflows?

For use with the NEBNext® Enzymatic Methyl-seq Kit (NEB #E7120), DNA should be sheared in one of the following buffers: 10 mM Tris-HCl pH 7.5 or pH 8.0, 1X TE (10 mM Tris-HCl pH 8.0, 1 mM EDTA), or low TE (10 mM Tris-HCl pH 8.0, 0.1 mM EDTA). We do not recommend shearing input DNA in 0.1X TE (1 mM Tris-HCl, 0.1 mM EDTA) or water.

For use with the NEBNext® Enzymatic Methyl-seq Conversion Module (NEB #E7125), the DNA must not be in a buffer that contains EDTA before proceeding with the Oxidation Reaction. Therefore, it is recommended that the DNA is sheared in 10 mM Tris pH 8.0. Alternatively, if shearing in 1X TE or low TE, a column or bead cleanup followed by elution in 10 mM Tris-HCl pH 8.0 or nuclease-free water is required.