Analysis of SOS induction at the population level by flow cytometry

Overnight cultures grown in MOPS-based medium supplemented with 0.4% glucose were diluted to an OD600nm of ~0.01 in the same medium. Cultures were grown for 5 hours to mid-log phase (OD600nm ~0.3) and diluted in phosphate-buffered saline (PBS) buffer before injection into an Attune NXT flow cytometer. In each experiment, 75,000 events were analyzed with a blue laser (488 nm) and a 530/30 emission filter for GFP and with a yellow laser (561 nm) and a 620/15 emission filter for Crimson (red fluorescence). Subsequent analyses were performed using FlowJo 10.4 and MATLAB (R2017b, MathWorks), assisted by custom-made scripts.

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