RNA-seq of Staphylococcus aureus human and bovine strains cultured in TSB and 50% milk media
RNA-seq of Staphylococcus aureus human and bovine strains cultured in TSB and 50% milk media
Overnight TSB Staphylococcus aureus cultures were standardised to OD600 0.1 in TSB and OD600 0.25 in 50% Milk/TSB cultures in 5ml volumes. All samples were incubated at 37°C; 180 rpm for 2h before pelleting at 4863xg 5 min. The pellet was suspended in 200 µl TE buffer and transferred into a 96-well plate and centrifuged. The supernatant was removed and the pellets immediately frozen on dry ice containing 100% ethanol. The frozen pellets were briefly stored at -80°C before shipment to Genewiz, UK for RNA extraction and paired-end RNA sequencing. RNA was extracted from Staphylococcus aureus using homogenization in the tissue Lyzer II (Qiagen) with 0.1 mm beads and beating at 30 Hz. The RNeasy Plus Mini Kit from Qiagen was used for RNA extractions. Libraries were prepared by GENEWIZ - Enrichment, cDNA synthesis, and adapter ligation