GSM5288835: S coelicolor mRNA day 1_rep2; Streptomyces coelicolor; RNA-Seq
cells were collected every 24 hours for 8 days after initial transfer to 1L cultures, quiagen RNAeasy kit was used. Cells were resuspended in RLT buffer with ten 3mm glass beads and then vortexed, followed by an additional equal volume of acid phenol chloroform. Samples were then vortexed and centrifuged at 15,000 rpm (10,000 x g) for 10 min at RT. The aqueous was isolated and diluted with an equal volume of 70 % (V/V) ethanol followed by purification with the RNAeasy column according to manufacturer's instructions. To remove gDNA contaminants an on column digestion was performed with 2 µL of TurboDNase. An Agilent 2100 Bioanalyzer was used to confirm the integrity of the RNA by assessing the 23s and 16s RNA bands. After ribozero treatment and library creation, sequencing was performed on an Illumina HiSeq 2500 system in high-output mode with 50 bp single-reads.