MND Buffer

1.2 m sorbitol

10 mm CaCl₂

100 mm NaCl

1 mm EDTA (pH 8.0)

50 mm Tris–HCl (pH 8.0)

0.075% NP-40

14 mm β-mercaptoethanol

5 mm spermidine

1% Protease Inhibitor Cocktail (Sigma-Aldrich P8215)

1 mm PMSF

Combine the first six ingredients filter-sterilized. (Use freshly prepared 2.4 m sorbitol stock solution.) Add β-mercaptoethanol, spermidine, Protease Inhibitor Cocktail, and PMSF immediately before use.

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