GSM5514342: S. maltophilia D457 bacterial culture collected at 0.6 OD600nm - Rep 4; Stenotrophomonas maltophilia; RNA-Seq
Twenty milliliters of each bacterial culture was spun down at 6,000 × g for 3 min at 4°C and immediately frozen on dry ice and stored at −80°C. RNA was isolated from cell pellets using the RNeasy minikit (Qiagen) following the manufacturer's instructions. To remove the remaining genomic DNA, total RNA samples were treated with DNase I (RNase-Free DNase Set [Qiagen]), and a second digestion was then performed following the Turbo DNA-free (Ambion) protocol. The RNA was purified and concentrated using RNeasy minikit columns. Finally, DNA contamination was checked by PCR with primers 27 and 48. Only RNAs containing no DNA contamination were used for further studies. Libraries were constructed with Truseq stranded total RNA Library and rRNA depleted with NebNext rRNA depletion kit. Sequencing was conducted by Illumina NovaSeq 6000 sequencing technology using a 150-bp paired-end format and 20 million total reads/sample, using three independent biological replicates of each sample