GSM8909615: P234_coreB_tissue; Homo sapiens; RNA-Seq
RNA from regions of interest was extracted from FFPE material using the Ambion Kit. The NEBNext Ultra II Directional RNA Library Prep Kit for Illumina (NEB #E7760S/L) was employed for fragmentation and cDNA synthesis. Depending on RNA quality, two fragmentation conditions were used: 10 minutes at 94°C for samples with RQN ≥ 4, and 5 minutes priming at 65°C for others. Following fragmentation, cDNA synthesis took place. Quality control was subsequently conducted using Qubit to ascertain the starting concentration. Sample preparation and hybridization capture were executed in line with the manufacturer's protocol for the SureSelectXT Low Input Target Enrichment System for Illumina Paired-End Sequencing Library (protocol v1.8, G9703-90000). For the hybridization process, Agilent SureSelectXT Human All Exon v7 Capture Library (5191-4006) baits were utilized. Clustering and sequencing using the NovaSeq6000 was performed according to manufacturer's protocols.