Recovery_120
Escherichia coli O17 str. K12a
Protocols: Escherichia coli was grown overnight in luria broth at 37°C with shaking at 200 rpm. The culture was back diluted to ~OD600 0.01 and allowed to grow to OD600 ~0.1. It was then treated with 1 μg/ml mitomycin C for 60 min. Cells were pelleted, washed with fresh media, resuspended to OD600 ~0.1 and then allowed to recover from damage treatment. Cultures were maintained in log phase (OD600 ~0.1 - 0.4). Samples were collected before addition of damage as control. Samples were also collected every 20 min for 3 hrs during recovery and then every 1 hr for 3 hrs after that. 1 ml of culture was pelleted, supernatant removed and stored at -80C. Escherichia coli was grown in luria broth at 37°C with shaking at 200 rpm. Escherichia coli was grown overnight in luria broth at 37°C with shaking at 200 rpm. The culture was back diluted to ~OD600 0.01 and allowed to grow to OD600 ~0.1. It was then treated with 1 μg/ml mitomycin C for 60 min. RNA was extracted using the Direct-zol™ RNA MiniPrep (Zymo, Cat no. R2052A81) and RNA Clean & Concentrator™-25 (Zymo, Cat no. R1018A82). RNA libraries were prepared using the TruSeq Stranded mRNA Library Preparation kit