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METHODS Maintenance and growth conditions of the organism. Escherichia coli ~1485 was maintained on slopes of tryptone soya agar (Oxoid) and grown in a basic minimal medium containing (per litre distilled water): 0.3 yo (w/v) DL-alanine as carbon source; 2 g (NH,),SO,; IOO ml 1-0 M-phosphate buffer pH 7-2; 5 ml mineral salts solution (10 g MgSO,. 7H,O, I g MnCl,. 4H,O, 0.05 g FeSO, and 0.1 g CaCl,/I distilled water). Cell counts. Culture samples (0.9 ml) were added to 0-1 ml of 40 yo (w/v) formaldehyde, agitated with a Whirlimix (Hook & Tucker Instruments Ltd., Croydon, Surrey) for 30 s, and the number of cells counted in a Helber bacterial counting chamber (Gallenkamp & Co., London) within 8 h. Synchronization of exponentially-growing cells. Starter cultures were grown from tryptone soya agar slopes in a shaking waterbath at 37 "C and used to inoculate a larger volume of prewarmed medium at a 1/200 dilution. The culture was force-aerated at I 1 air/min/l at 37 "C for 17 to 18 h when cells were in the late-exponential phase of growth at between 5 x IO* and 109 cells/ml. The procedure for continuous-flow size selection synchrony has been given in detail by Lloyd et al. (1975). The MSE continuous flow rotor and all attached tubing was autoclaved (103 kN m-, for 20 min) before use and left overnight at 37 "C. The thermostat on the MSE 18 high speed

Preparation of Synchronous Cultures of Escherichia coli by Continuous-flow Size Selection