GSM4307176: 3xdelta-relQ-30min-ohne-AHT-e_S9_R1_001; Staphylococcus aureus; RNA-Seq
Pellet was solved in Trizol. Strains were lysed using 0.5mm circonia silicia beads. Chloroform was used seperate RNA from DNA and proteins. RNA extraction was performed following the protocol by Amp Tech ExpressArt® RNA ready RNA was examined by a capillary electrophoresis on a Shimadzu MultiNA microchip followed by rRNA depletion using Ribo-Zero rRNA removel Kit from Illumina. RNA was converted to cDNA by fragmenting RNA samples by ultrasound and ligating an oligonucleotide adapter to the 3'end of the RNA. Using M-MLV reverse transcriptase first strand cDNA was created using 3' adapter as primer. The 5'Illumina TruSeq sequencing adapter was ligated to the 3'end of the purified (Agencourt AMPure XP kit) cDNA and PCR was performed. Samples were pooled in equimolar amounts and fractionated in a size range of 200-500 bp using a preparative agarose gel and Illumina sequencing was performed using 75bp reads.