P-EMBL-150 - P-EMBL-150
Yeast cells were streaked out on YPD agar plates. After growth for 2 days at 30°C an overnight culture (20 ml) was inoculated in YPD medium (160 rpm; 30°C). The main culture (lactate medium; 30 ml) was inoculated with the overnight culture to an OD600 of 0.2. Cells were grown to an OD600 of 1.0 (160 rpm; 30°C), harvested, washed once with sterile distilled water, snap frozen in liquid nitrogen and stored at -80°C.
YPD: 1% (w/v) bacto yeast extract; 2% (w/v) bacto peptone; 2% (w/v) glucose.
Lactate medium: 0.3% (w/v) yeast extract; 2% (v/v) lactic acid; 0.1% (w/v) glucose; 0.1% (w/v) KH2PO4; 0.1% (w/v) NH4Cl; 0.05% (w/v) CaCl2x2H2O; 0.05% (w/v) NaCl; 0.06% (w/v) MgCl2xH2O; 0.3 ml of a 1% (w/v) FeCl3 solution per 1 l medium; pH 5.5 (KOH).
media, min temperature, start time