FAQ: Can I fragment my gDNA sample with NEBNext UltraShear® in buffers other than 1X TE (10mM Tris pH 8.0, 0.1mM EDTA)?

gDNA samples can be fragmented with NEBNext UltraShear® with common gDNA extraction kits’ elution buffers, e.g. Monarch® DNA Elution Buffer, Monarch® gDNA Elution Buffer, Monarch® gDNA Elution Buffer II, Buffer EB and Buffer AE, in place of 1X TE pH 8.0. However, the fragmentation for NEBNext UltraShear is slower with gDNA samples fragmented in these elution buffers. Fragmentation conditions would need to be optimized and fragmentation may need to be incubated for extended times at 37˚C or 45 ˚C compared to gDNA samples in 1X TE pH 8.0.

Monarch® DNA Elution Buffer (NEB #T1016): 10 mM Tris, 0.1 mM EDTA pH 8.5
Monarch® gDNA Elution Buffer (NEB #T3016): 10 mM Tris, 0.1 mM EDTA pH 9.0
Monarch® gDNA Elution Buffer II (NEB #T3056) or Buffer AE: 10 mM Tris-HCl, 0.5 mM EDTA pH 9.0
Qiagen Buffer EB: 10 mM Tris-HCl pH 8.5