Cell cycle mRNA half-life profiling.

mRNA half-life profiling in the bacterium Caulobacter crescentus was performed in synchronized cells collected from different stages of the cell cycle including swarmer cells, stalk cells (45 min post synchrony), and predivisional cells (90 min post synchrony). For each cell population, transcription was disrupted by the antibiotic rifampicin, and RNA samples were collected at different time points to measure the mRNA half-lives. Overall design: mRNA half-life profiling in the bacterium Caulobacter crescentus was performed in synchronized cells collected from different stages of the cell cycle including swarmer cells, stalk cells (45 min post synchrony), and predivisional cells (90 min post synchrony). For each cell population, transcription was disrupted by the antibiotic rifampicin, RNA samples were collected. All RNA measurements were performed on cells grown to mid-log in M2G minimal growth medium. Total RNA samples were collected just prior to rifampicin addition, and after 1, 3, or 9 minutes incubation with 100ug/mL rifampicin. Each total RNA sample was depleted of rRNA using the ribozero kit (Illumina) for gram negative bacteria, and subjected to total RNA-seq libraries as in (Ingolia et al. Science 2009). tRNA was left in each sample as an internal control for stable RNA.