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04 Add slowly Insert the needle and let the water run down the inside wall of the vial

harvested cells were first re-suspended in chilled lysis buffer (20 mM Tris [pH 7.9], 0.5 M NaCl, 5 mM imidazole, 10% glycerol, 1 mM DTT, 1 mM PMSF) and the lysis mixture was sonicated on ice