GSM9283787: grtKmutant; Burkholderia glumae; RNA-Seq
Total RNA was isolated from B. glumae wild-type BGR1, the grtK mutant (BGR1 grtK::Tn3-gus24) and the grtR mutant (BGR1 grtR::Tn3-gus54), using the RNeasy Mini Kit (Qiagen, Venlo, Netherland) according to the manufacturer's instructions. To remove genomic DNA completely, the isolated RNA was treated with DNaseI (Ambion, Austin, TX, USA) for 1 h at 37 °C according to the supplier's instructions. The quantity and quality of the total RNA were evaluated using RNA electropherograms (Agilent 2100 Bioanalyzer, Santa Clara, CA) and by assessing the RNA integrity number (RIN). The NEBNext rRNA Depletion Kit (Bacteria) (New England BioLabs, UK) was used for ribosomal RNA depletion according to the manufacturer's instructions. Libraries for Illumina sequencing were made with the TruSeq stranded mRNA library prep (Illumina, USA) following the manufacturer's protocol.