title: BRIC_PolyAminus_Hela_15min
organism: Homo sapiens
description: HeLa cells were incubated at 37oC in the presence of 150uM 5'-bromo-uridine (BrU) (Wako, Osaka, Japan) for 24 h in a humidified incubator with 5% CO2. After replacing BrU-containing medium with BrU-free medium, cells were harvested at indicated time points. Total RNA was isolated using RNAiso Plus (TaKaRa). Twelve micrograms of BrU-labeled total RNA were denatured by heating at 80oC for 1 min and then added to anti-BrdU mAb-conjugated beads containing 2ug of anti-BrdU mAb (clone 2B1, MBL). The mixture was incubated at room temperature for 1 h with rotation. Beads were washed four times with 0.1% BSA in PBS. ISOGEN LS (Nippon Gene, Tokyo, Japan) was added, followed by RNA isolation, according to the manufacturer's instructions. The isolated RNA was used for deep sequencing using the mRNA-seq Sample Preparation Kit using the same protocol as RNA-seq. Data processing was conducted by the identical procedures as the RNA-seq method above. For BRIC-seq data without transfection, we used 13 time points to calculate half-life: 15 min
attributes.tag: sample_name
attributes.value: BRIC_PolyAminus_Hela_15min
attributes.tag: cell_line
attributes.value: Hela