E-MTAB-9696
Asymmetric chromosome segregation and cell division in DNA damage-induced bacterial filaments
We performed RNA seq analysis on Escherichia coli MG1655 recovering from DNA damage induced by mitomycin C 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. 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. Cultures were maintained in log phase (OD600 ~0.1 - 0.4) throughout the experiment. 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. Libraries were sequenced using Illumina MiSeq sequencing platform. Samples were collected from 2 independent experiments.
We performed RNA seq analysis on Escherichia coli MG1655 recovering from DNA damage induced by mitomycin C 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. 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. Cultures were maintained in log phase (OD600 ~0.1 - 0.4) throughout the experiment. 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. Libraries were sequenced using Illumina MiSeq sequencing platform. Samples were collected from 2 independent experiments.