GSM8324341: P-shR, 24hours, rep3; Homo sapiens; RNA-Seq
Cells were collected using trypsinization and washed with ice-cold PBS and centrifuged at 1500 rpm for 5 minutes. Total RNA was extracted from cell pellet using Qiagen RNeasy Plus kit. Total RNA examples were checked for quality on Agilent Bioanalyzer 2100 and quantity with Qubit fluorometer. 1.3 ug of total RNA was used for the construction of sequencing libraries The NEBNext Ultra II RNA Library Prep Kit for Illumina was used to prepare sequencing libraries. The Kit procedure was performed without modifications. This procedure includes rRNA depletion, remaining RNA purification and fragmentation, cDNA synthesis, end preparation, Illumina adapter ligation, library PCR amplification and validation.