GSM8766495: strain 1c, biological replicate 3; Candidozyma auris; RNA-Seq
Cells were collected by centrifugation, supernatants removed, and the cell pellets stored at -80degC until RNA isolation. Cell cultures were grown in triplicate. Cell pellets were thawed and resuspended in 100 µl FE Buffer (98% formamide, 0.01 M EDTA) at room temperature. Fifty microliters of 1 mm RNase-free glass beads was added, and the cells were disrupted in a vortex disruptor for 5 min at room temperature followed by snap-cooling on ice. The cell lysate was centrifuged to remove cell debris, and the crude RNA extract collected from the supernatant. The extract was DNase-treated for 30 min and precipitated by isopropanol precipitation. RNA integrity was confirmed through agarose gel electrophoresis using the bleach gel method, and concentrations were approximated by Nanodrop. RNA sequencing was performed using Illumina NextSeq for stranded mRNA. Libraries were prepared with paired-end adapters using Illumina chemistries per manufacturer's instructions, with read lengths of approximately 150 bp with at least 50 million raw reads per sample.