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The cel ls were pelleted after 20 hours by centrifugation using a Fiberlite F9-6×1000 rotor in a Sorvall LYNX 6000 Superspeed centrifuge at 2,000 g for 20 minutes, re-suspended in 40 ml lysis buffer containing 50 mM Tris, pH 7.5, 150 mM NaCl, 20 mM imidazole, 20 % glycerol, 2 mM TCEP, 250 units of Benzonase® Nuclease and bacterial Protease Inhibitor Cocktail (Amresco