Total RNA was extracted from PC3 and DU145 cells treated with 2.5 uM pentamidine or vehicle using the TRIzol reagent (Invitrogen, 15596018) and mRNA was enriched by Oligo(dT) beads. The enriched mRNA was fragmented into short fragments using the fragmentation buffer and reverse transcribed into cDNA with random primers. Second-strand cDNA fragments were synthesized by DNA polymerase I, RNase H, dNTP and buffer. Then these cDNA fragments were purified with QiaQuick PCR extraction kit, end repaired, poly(A) added, and ligated to Illumina sequencing adapters. The ligation products were then size selected by agarose gel electrophoresis, PCR amplified, and sequenced using Illumina Novaseq 6000 PE150.
GEO Accession
GSM3887766