GSM3040168: KS20; Homo sapiens; RNA-Seq
Surgical specimens were transferred immediately from the operating room to the surgical pathology lab where a section of full-thickness sigmoid colon was obtained away from sites of active inflammation and stored in RNAlater. RNA was extracted from tissue samples using trizol reagent followed by purification with the Qiagen RNeasy Mini Kit. The cDNA libraries were prepared using the NEXTflex® Rapid Directional qRNA-Seq™ Kit (24 barcodes) (BioO Scientific) as per the manufacturer's instructions. The final product was assessed for its size distribution using the Bioanalyzer DNA High Sensitivity Kit (Agilent Technologies) and for its concentration using Kapa library quantification kit (Kapa Biosystems). Six libraries were pooled per HiSeq lane, followed by on-board cluster generation on a Rapid Run single-end flow cell and subsequent 50 cycles sequencing (v3 sequencing kit) according to the manufacturer's instructions (HiSeq 2500, Illumina).