GSM9422160: PDAC_ex_vivo_slice, OT138, Selenite_5uM_(24-48h), 48h; Homo sapiens; RNA-Seq
Total RNA was isolated from 10 um DNase-treated FFPE sections using the Maxwell RSC FFPE RNA Kit (Promega AS1440) according to the manufacturer. RNA quantity/quality was assessed using Qubit 4.0 with the RNA HS Assay Kit (Thermo Fisher Q32852). Up to 50 ng total RNA per sample was used for library preparation with the SMARTer Total RNA-Seq Kit v2.5 Pico Input Mammalian (Takara 634411). RNA was thermally fragmented (94 C, 3 min) and reverse-transcribed with random N6 hexamers. Illumina adapters and sample barcodes were introduced during amplification; ribosomal cDNA was depleted by RNase H with mammalian R-probes; remaining fragments were re-amplified with universal adapter primers. Libraries were quantified (Qubit DNA HS, Q33230), normalized, pooled and sequenced on an Illumina NextSeq 500/550 Mid Output v2.5 (150 cycles) to generate 2 x 75 bp paired-end reads.