sentence: Approximately 100OD of cells grown in YPD were harvested by centrifugation at 3000 g and washed with LB buffer ( 50mM Tris / HCl [ pH7.4 ] , 200mM NaCl , 1mM EDTA , 2mM phenylmethylsulfonyl fluoride ( PMSF ) and protease inhibitor cocktail ) .