GSM8615836: Elim_WT_lactate_without_W_1; Eubacterium limosum; RNA-Seq
E. limosum cells were grown in 50 mL serum bottles under glucose and lactate conditions, with and without 100 nM W added. At mid-log phase (OD600 = 0.3), 50 mL of cells were spun down for 15 minutes at 15,000g at 4 °C. The cells were resuspended in 0.5 mL of the supernatant and 2 mL RNAprotect Bacteria Reagent (Qiagen). The Qiagen RNeasy Mini Kit was used as suggested with an added lysozyme treatment and bead beater (model and parameters) was used to extract RNA. An additional on column DNase I treatment was performed as suggested with the kit. The RNA quality was assessed using A260/A280 on a Nanodrop and a Bioanalyzer RIN>7 (as suggested by Novogene). 1 µg of RNA was used for library preparation. Ribosomal RNA was removed from total RNA, followed by ethanol precipitation. Sequencing libraries were generated using NEBNext Ultra RNA Library Prep Kit for Illumina following manufacturer recommendations and index codes were added.