GSM7927851: CFPAC-1 cell, CGN siRNA; Homo sapiens; RNA-Seq
Total RNA was isolated from CGN knockdown (CGN siRNA) or control (NC) CFPAC-1 cells using Trizol reagent (Company, City, Country). RNA quality was evaluated on an Agilent 2100 Bioanalyzer (Agilent Technologies, Palo Alto, CA, USA) and checked by RNase-free agarose gel electrophoresis. After total RNA was extracted, eukaryotic mRNA was enriched using oligo(dT) beads. The enriched mRNA was then fragmented into short fragments using fragmentation buffer and reverse transcribed into cDNA using the NEBNext Ultra RNA Library Prep Kit for Illumina (CAT# 7530, New England Biolabs). The purified double-stranded cDNA fragments were end repaired, added with A base, and ligated to Illumina sequencing adapters. The ligation reaction was purified using AMPure XP Beads (1.0X). Ligated fragments were size selected by agarose gel electrophoresis and amplified by polymerase chain reaction (PCR). The resulting cDNA library was sequenced by Gene Denovo Biotechnology Co. (Guangzhou, China) using Illumina Novaseq6000.