GSM3463545: wild type, no 3-OH-C10-HSL, replicate 1; Methylomonas sp. LW13; RNA-Seq
Cell pellets were lysed by bead beating with 0.1-mm zirconia-silica beads (Biospec Products) in 1 mL TRIzol (Thermo Fisher Scientific). The lysate was then separated according to the TRIzol instructions, and subsequently 1.5 volumes of 100% ethanol was added to the aqueous phase of the extract which was then used as the input for an RNeasy RNA isolation kit (Qiagen). The resulting elution was then digested using Ambion DNase I (Thermo Fisher Scientific) for 30 minutes at 37ºC before being re-purified via RNeasy RNA isolation kit with the addition of an on-column DNase (Qiagen) digestion step for 15 minutes at room temperature. The resulting purified RNA was checked for DNA contamination by PCR using the universal 16S primers 27F and 1492R. cDNA library preparation and RNA sequencing were performed by GENEWIZ using Illumina HiSeq 2x150 (pair ended) reads.