GSM9521138: B4G12 cells, siControl3; Homo sapiens; RNA-Seq
Total RNA was extracted using RNeasy Micro Kit (Cat# 74004, Qiagen)following the manufacturer's instructions and checked for a RIN number to inspect RNA integrity by an Agilent 2100 Bioanalyze (Agilent technologies, Santa Clara, CA, US).Qualified total RNA was further purified by RNAClean XP Kit (Cat A63987, Beckman Coulter,Inc.Kraemer Boulevard Brea, CA,USA)and RNase-Free DNase Set  (Cat#79254, QIAGEN, GmBH, Germany) The purified Total RNA was subjected to a series of steps including mRNA isolation, fragmentation, first-strand cDNA synthesis, second-strand cDNA synthesis, end repair, 3'-end adenylation, adapter ligation, and PCR enrichment to complete the construction of the sequencing sample library. The concentration of the constructed library was detected using a Qubit® 2.0 Fluorometer, and the library fragment size was verified by an Agilent 4200 TapeStation. Sequencing was performed on an Illumina NovaSeq 6000 platform with a PE150 (Paired-End 150 bp) sequencing mode.  Approximately 8 Gb of clean data per sample. The proportion of bases with a quality score higher than 20 (Q20) in each read direction shall be no less than 90%.