GSM5618002: ER00573, 5 hours, Replicate 1 [MRSA-37]; Staphylococcus aureus; RNA-Seq
Samples were bead beated using FastPrep and spun down. Upper phase was collected and 200µL chloroform was added. 0.5mL isopropanol was added to aqueous phase to precipitate RNA. RNA was washed with 70% ethanol, air dried, and resuspended in RNase free water. 2500ng of RNA was DNase treated (TURBO DNA-freeTM kit, Invitrogen Ambion). For each sample, 250ng of RNA was first depleted of bacterial rRNAs using enzymatic ribodepletion with the Illumina Ribo-Zero Plus rRNA Depletion Kit (Illumina, Catalog no. 20037135) according to the manufacturers protocol. RNA-Seq libraries were then prepared using the Illumina TruSeq Stranded Total RNA Library Prep kit using the depleted rRNA material as input and following the manufacturer's protocol.