title: GSM8792796: Tubular_Adenoma_6; Homo sapiens; RNA-Seq
library_name: GSM8792796
library_construction_protocol: Three sequential sections, each 3µm thick, from formalin-fixed paraffin-embedded (FFPE) tissue biopsies from the cases mentioned in Table 4 were subjected to RNA extraction using the Recover All kit according to the manufacturer's instructions (ThermoFisher Scientific, USA). Genomic DNA was removed from all RNA samples by treating the extracted RNA with the TURBO DNAase-free™ Kit (Invitrogen, Carlsbad, CA, USA). A total of 100 ng of extracted and Turbo DNase treated RNAs, was utilized for whole transcriptome sequencing from 29 endoscopic biopsy samples. This was accomplished utilizing a targeted AmpliSeqTranscriptome Panel, meticulously designed to target over 21,000 human RNA transcripts through a high-throughput multiplexed approach on the Ion S5 XL Semiconductor Sequencer (Thermo Fisher Scientific, USA) equipped with the Ion 540 Chip (Life Technologies, Carlsbad, CA, USA). In summary, the total RNA underwent processing using the SuperScript VILO cDNA synthesis kit (Invitrogen; 11754050), followed by amplification of the resulting DNA using the lon AmeliSeq™ Transcriptome Human Gene Expression Kit (Thermo Fisher Scientific; A26325) for library preparation. Subsequently, the libraries underwent purification using Agencourt AM-Rute XP Beads (Beckman Coulter, Indianapolis, IN, USA) and quantification employing the lon Library TaqMan™ Quantitation Kit (Applied Biosystems, Waltham, MA, USA). The libraries were diluted to -100 pM and pooled by combining equal volumes of each barcoded library. The resulting template libraries were then sequenced on the Ion S5 XL Semiconductor sequencer using the Ion 540 Chip (Life Technologies Corporation, Carlsbad, CA) prepared on fully automated Ion Chef System (Thermo Fisher Scientific).