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298 Serebriiskii et al.Table 1Concentrations of Antibiotics and Other Supplements Used in Agar PlatesFinalconcentrationAntibiotic Stock solution in plates Abbreviation aAmpicillin 100 mg/mL in H 2O 100 µg/mL AChloramphenicol 30 mg/mL in ethanol 15 µg/mL CKanamycin 10 mg/mL in H 2O 10 µg/mL KTetracycline 10 mg/mL in 80% ethanol 5 µg/mL TStreptomycin 10 mg/mL in H 2O 10 µg/mL SIPTG 1 M in H 2O Up to 50 µM I3-Aminotriazole 1 M in H 2O 1–40 mM ATa Media description consists of the name of basic medium, followed by abbreviations forantibiotics and inducers, followed by concentration of aminotriazole. For example, LB_AC indicatesLB plates with ampicillin and chloramphenicol; NM_ACSI_5AT is NM medium with ampicillin,chloramphenicol, streptomycin, IPTG, and 5 mM 3-Aminotriazole.418 mL ddH 2O50 mL 10X M9 salts (Miller recipe [16,17]:for 1 L, add 60 g Na 2HPO 4, 30 gKH 2PO 4, 5 g NaCl, and 10 g NH 4Cl,autoclave)10 mL 20% Glucose5 mL 20 mM Adenine HCl15 mL Amino acid mixture (see Subheading 2.1.)0.5 mL 1 M MgSO 40.5 mL 10 mg/mL Thiamine0.5 mL 10 mM ZnSO 40.5 mL 100 mM CaCl 2(always add this last).2. NM agar plates: for 500 mL of plates, autoclave 418 mL ddH 2O with 7.5 g ofbacto-agar and a stir bar, allow agar to cool to 65–70°C and then add the same82 mL of basic components as for the liquid NM medium (premixed and filtered).3. Liquid Luria-Bertani (LB) medium: add 5 g bacto-tryptone, 2.5 g yeast extract, and5 g NaCl to 400 mL dH 2O, adjust pH to 7.5 with NaOH, adjust volume to 500 mL,autoclave 15 min.4. LB agar plates: add 7.5 bacto-agar to liquid LB media before autoclaving.Add antibiotics, IPTG, and 3-AT to media as needed (see text for details).When adding antibiotics, ensure temperature of autoclaved media is less than70°C before addition.