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25 Two days after hatching 100 μ l/4 ml agar of 5 × metabolite in the M9 salt buffer (3 mg/ml KH 2 PO 4 , 6 mg/ml Na 2 HPO 4 , 5 mg/ml NaCl, 1 mM MgSO 4 ) or M9 (with appropriate amounts of EtAc) is supplemented to an NGM plate to refresh the metabolite without the need for transfer
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