GSM7072979: biol rep 1, S.aureus+ tonsillar cells , test, 1h; Staphylococcus aureus; RNA-Seq
After 1 and 3h post-infection, the media was aspirated, and the host cells were washed twice with fresh media to remove unbound bacteria. The host cells were then trypsinized and lysed with Triton-X. The released bacteria were then collected from three technical replicates and pooled together for RNA extraction. Plate enumeration, adhesion assay and cytotoxicity assay were also performed. RNA was harvested using Rneasy mini plus kit (Qiagen). 1.2 ug of total RNA was used for the construction of sequencing libraries. Prior to RNA libraries for RNA-seq, depletion of bacterial rRNA was performed with the RiboCop rRNA depletion kit for Gram-Positive Bacteria (G+), according to the manufacturer's protocol. In total 15 samples were processed for libraries construction using Lexogen's CORALLTM Total RNA-Seq Kit with RiboCop following  he manufacturer's protocols.