GSM4626712: MCF7_ctl_r2; Homo sapiens; OTHER
Cells were labeled with 2 mM Bromouridine (Sigma Aldrich) for 30 min, total RNA was isolated by using TRIzol reagent (Invitrogen), and Bru-labeled RNA was isolated from the total RNA by incubation with anti-BrdU antibodies (BD Biosciences) conjugated to magnetic beads (Dynabeads, Goat anti-Mouse IgG; Invitrogen) under gentle agitation at room temperature for 1 h. For more detail, see Paulsen et. al 2013 (Coordinated regulation of synthesis and stability of RNA during the acute TNF-induced proinflammatory response. Proc. Natl. Acad. Sci. U.S.A. 2013; 110:2240–2245). cDNA libraries were made from the BrU-labeled RNA using the Illumina TruSeq library kit and sequenced using Illumina HiSeq sequencers at the University of Michigan DNA Sequencing Core with 52 bp single-end reads (on average 50 million reads per sample including 70% unique reads). Sequencing and read mapping (unique reads only) were carried out as described using the human genome reference assembly hg19/GRCh37 (Paulsen et al., 2014).
GEO Accession
GSM4626712