SAMD00010796
External Id
Laboratory of Functional Genomics, Department of Medical Genome Sciences, Graduate School of Frontier Sciences, The University of Tokyo
INSDC center name
2015-03-16T00:00:00Z
INSDC first public
2015-03-15T20:20:40Z
INSDC last update
DRS012771
INSDC secondary accession
live
INSDC status
Generic
NCBI submission model
Generic.1.0
NCBI submission package
DRS012771
SRA accession
Homo sapiens
http://purl.obolibrary.org/obo/NCBITaxon_9606
organism
This sample is total RNA that was treated BRIC in the control HeLa cells.It was harvested in 465min time point.BRIC was performed as previously described. In brief, cells were incubated at 37Â°C in the presence of 150μM 5'-bromo-uridine (BrU) (Wako) for 24 h in a humidified incubator with 5% CO2. At indicated time points after replacing BrU- containing medium with BrU-free medium, cells were harvested. Total RNAs were isolated using RNAiso Plus (TaKaRa). Twelve micrograms of BrU-labeled total RNA was denatured by heating at 80Â°C for 1 min and then added to the anti-BrdU mAb- conjugated beads containing 2μg 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) was added, followed by RNA isolation in accordance with the manufacturer’s instructions. The isolated RNA was used for deep sequencing using mRNA-seq Sample Preparation Kit using the same protocol as RNA-seq. For STAU1 knockdown data, 11 time-points were taken, 0mins, 15 mins, 45 mins, 75mins, 105mins,165mins, 225mins, 345mins, 465mins, 585mins and 705mins.HeLa cells were grown in Dulbecco’s Modified Eagle’s Medium (DMEM) supplemented with 10% Fetal Bovine Serum and antibiotics at 37oC at 5% CO2 in a humidified incubator.siRNAs were transfected using Lipofectamine RNAiMAX (Invitrogen), according to the instructions from the manufacturer. Following the siRNA duplex transfection (final concentration 10nM), cells were harvested for 72h after the transfection.
sample comment
DRS012771
sample name
BRIC_siCTRL_465min_2
title