GSM2811258: Saureus_HG003_RNAseq; Staphylococcus aureus; ssRNA-seq
Cultures were centrifuged; the pellets were frozen in liquid nitrogen and stored at -80°C until RNA extraction. Cells pellets were resuspended into 400 μL of Lysis buffer (4 M guanidine thiocyanate, 25 mM sodium acetate pH 5.2, 5 g/L N-laurylsarcosinate), transferred into FastPrep tubes containing 0.6 g of glass beads (G4649, Sigma-Aldrich) and 400 µL of acid phenol:chloroform:IAA (25:24:1). Bacteria were mechanically lysed by using the Fastprep apparatus (MP Biomedicals) with 3 cycles of 45 s at speed 6.0 separated by incubation on ice during 5 min. After lysis, tubes were centrifuged 15 min at 17,900 g at 4 °C. The aqueous phase was acid phenol extracted, isopropanol precipitated and the pellet resuspended in RNase-free water. Strand-specific RNA-seq libraries were generated with the TruSeq RNA-Seq Library Preparation Kit (Illumina), according to the manufacturer's instructions.
GEO Accession
GSM2811258