2 protocols
Total RNA was isolated from E. coli cells, grown overnight in presence of test compounds on a rotator shaker (150 rpm) at 37degrees C using Qiagen RNeasy Mini Kit column (Qiagen, CA, USA).
E. coli MG1655 was grown overnight in 50 ml of Luria Bertani (LB) medium on a rotatory shaker (150 rpm) at 37degrees C. Next day, the culture was diluted using saline (0.85%) to ~500 cfu/ml. Fresh LB (5 ml) was inoculated with 100ul of the diluted cell suspension and grown in presence of the test compounds (Plumbagin/Juglone/Menadione; 50uM) and solvent control (DMSO; 0.75%), separately, till the cell density reached to ~1 billion cfu/ml .