Fitness profiling using three different growth media (YES, EMM, and  EMM+K)
Frozen Bioneer version 1.0 haploid library in 96-well plate format (catalog   number M-1030H; received on April 24, 2008) was thawed at room temperature and five-  microliter portions of the glycerol stock were aspirated from the bottom of the 96-well   plates and transferred to deep well plates containing YES agar medium supplemented   with 150 mg/l G418 and 100 mg/l carbenicillin. After two days of incubation at 30°C,   liquid YES medium supplemented with G418 was added and the strains were grown for   two more days in a shaker. The liquid cultures were pooled together and briefly   centrifuged. The cell pellets were resuspended to a concentration of 15.0 OD600 units per  ml with fresh liquid YES medium containing Hogness Freezing Medium. The cell   suspension was aliquoted into 1.5 ml microtubes at 0.5 ml per tube (7.5 OD600 units) and  frozen at -80°C. The recipe for 10× Hogness Freezing Medium stock was: 360 mM   K2HPO4, 132 mM KH2PO4, 17 mM sodium citrate, 4 mM MgSO4, 68 mM (NH4)2SO4,   44% Glycerol (Werner et al. 1997). The 10x freezing medium was mixed with YES   medium at a 1:9 ratio before use.