GSM3900853: MCF-7_rep1_input; Homo sapiens; OTHER
Generally, with harvesting cells as the first step, the culture medium was sucked away with a pipette, and the cells were washed twice with proper volume of 1×PBS buffer for each plate. Then the cells were digested with trypsin-EDTA solution containing 0.25% trypsin and 0.02% EDTA in 1×PBS buffer, and centrifuged at 1000 g for 3 min. Subsequently, total RNA was isolated from cell lines with TRIzol reagent (Invitrogen) according to the manufacturer's instruction by isopropanol precipitation. For poly(A)+ RNA purification, mRNA was extracted by two successive rounds of poly(A)+ purification using Oligo d(T)25 Magnetic Beads (NEB). The untreated and pyridine borane treated samples were performed with NEBNext® Ultra™ II Directional RNA Library Prep Kit for Illumina® following its manufacturer's instruction in parallel for Illumina sequencing.
GEO Accession
GSM3900853