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'''2-step transformation procedure for ''Bacillus subtilis'' 168''' *Cells are grown O/N at 37 °C in [[supplemented medium]] contains [[Spizizen's salts]] (without antibiotics) *Dilute cells 10X in fresh medium (10 ml) and grow for 3 h at 37 °C, shaking *Make agar plates *Cells are then diluted 1:1 in [[starvation medium]](prewarmed, no antibiotic) *After 2 hours, approx. 1 μg DNA (BRB689; Cmr ''amyQ+'') is added to 100 μl cells in a 2 ml tube, and the mixture is incubated for 30 min at 37 °C (200 rpm) *Dilute 3-fold in prewarmed [[TY medium]] and plate after another 45 min (37 °C;200 rpm) *Dilutions (in starvation medium) for plating are: * 10^0 and 10^-1 on selective media * 10^-5 on non-selective plates (viable count)

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