GSM3446285: DAP-S Sample_1-A (small RNA); Staphylococcus aureus; ncRNA-Seq
A single colony was used to inoculate 7 ml of BHI liquid culture and incubated at 37 °C  for 12h. Total RNA was extracted with TRIzol reagent (Invitrogen) according to the manufacturer's protocol. Sequencing libraries were prepared using the Illumina mRNA-seq sample preparation kit following  the supplier's instructions except that total RNA was not fragmented and double-stranded cDNA was size-selected (100 to 400 bp) to maximize the recovery of small size RNAs. The prepared libraries were evaluated with the High sensitivity D1000 screen Tape (Agilent Tape Station 2200). The indexed libraries were quantified in triplicates with the ABI7900 qPCR instrument using the KAPA Library Quantification Kit, according to the manufacturer's protocol (KapaBiosystems, Woburn, MA, USA). Five μL of the pooled library at a final concentration of 4 nM were used for sequencing using Illumina Miseq with a A single end stranded library with reads of 50bp. After sequence data generation, raw reads were processed using FastQC v0.11.2 to assess data quality. Reads were then trimmed using Trimmomatic v.0.33.2 to remove sequencing adapters for Single End reads, requiring a minimum base quality of 15 (Phred scale) and a minimum read length of 15 nucleotides. Only trimmed reads were included into downstream analysis.
GEO Accession
GSM3446285