GSM5596002: Bacteria (GB00112) exposed to cells for 24hr; Streptococcus agalactiae; RNA-Seq
RNA samples were collected from bacterial culture in liquid medium by adding two volumes of RNAprotect Bacteria Reagent (Qiagen) or from bacteria inside macrophages by washing the cells twice with PBS then adding 1 mL RNAprotect Bacteria Reagent directly to the cells. RNA was extracted using the RNeasy minikit (Qiagen) using enzymatic lysis, Proteinase K digestion, and mechanical disruption. Residual genomic DNA was removed using the Turbo DNA-free kit (Ambion). Illumina TruSeq Stranded mRNA Library Prep Kit was used. The protocol was modified to skip the oligo-dT bead step and proceeded directly to fragmentation and first strand synthesis