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If you see particles floating after 10 minutes of gentle mixing, something went wrong

It's best to get treated for B12 deficiency anemia and avoid any long-term problems

Cells were directly lysed in Laemmli buffer (60 mM Tris-HCl (pH 6.8), 2% (w/v) sodium dodecyl sulfate (SDS), 10% (v/v) glycerol, and 0.02% (w/v) bromophenol blue), then sonicated and denatured by boiling at 95 for 5 minutes