CdnLDD60+G_rep2
For 24 hour starved and 60 minute recovery samples, cultures of 3 independent colonies were inoculated into 6 mL M2G and grown overnight. Once cultures were at an OD600 of 0.4  0.6, cultures were prepared for carbon starvation as described above except after final wash, cells were resuspended in 6 mL M2 and incubated at 30C shaking at 225 rpm for 24 hours. After 24 hours, 2 mL of each culture was removed and stabilized using RNAprotect as described above and flash frozen in liquid nitrogen. To the remaining 4 mL of culture, 40 L of 20% glucose solution was added, and cultures were incubated for 60 minutes. Recovery samples were stabilized with RNAprotect and flash frozen in liquid nitrogen. All samples were processed at SeqCenter (Pittsburg,PA). There, samples were DNAse treated with Invitrogen DNAse (RNAse free). Library preparation was performed using Illuminas Stranded Total RNA Prep Ligation with Ribo-Zero Plus kit and 10bp IDT for Illumina indices and Caulobacter-specific rRNA depletion probes. Sequencing was done on a NextSeq2000 giving 2x51bp reads.