GSM9888182: HPMEC + Ctrlsh-1; Homo sapiens; miRNA-Seq
Total RNA were isolated from cells harvested in Trizol using the phenol-chloroform extraction protocol. 1ug of total RNA were used to prepare miRNA library. Total RNA quality and integrity were assessed by NanoDrop spectrophotometry and Agilent Bioanalyzer/Fragment Analyzer analysis. Samples with RNA integrity number (RIN) ≥7 were used for library preparation. Small RNA libraries were generated using the Qiagen miRNA Library Prep Kit (Cat# 331505) according to the manufacturer's instructions. Briefly, 3′ and 5′ adapters were ligated to miRNAs, followed by cDNA synthesis with unique molecular identifier (UMI) assignment, PCR amplification, and library purification. Library quality was assessed using an Agilent TapeStation and quantified by qPCR. Libraries were sequenced on an Illumina NovaSeq X platform using 100-bp paired-end reads, targeting 10–25 million reads per sample.