GSM3874638
GSM3874638: CL5_4h; Homo sapiens; RNA-Seq
Cells were harvested at 1, 2, 4, 8, 12, and 24 hours after addition of 4SU. Harvesting was performed by dislodging cells off the plate during two washes with cold PBS followed by spinning at 1,000G for 5 minutes at 4°C. Cell pellets were resuspended in 1mL TRI-Reagent and extracted according to manufacturer instructions. Extracted samples were spiked with Drosophila melanogaster and Saccharomyces Cerevisiae RNA. For timepoints, 4SU labeled RNA was purified as described in detail in the published methods. In addition, an unpurified subset of the 24 hour RNA was taken as the “Steady State” sample. Purified 4SU labeled RNA or unpurified total RNA from the 24-hour time point was used to prepare sequencing libraries using the TruSeq Stranded mRNA Sample Preparation Kit (Illumina), according to manufacturer's instructions. Adapter-ligated fragments were enriched with 14x PCR cycles. ~16-22 samples were multiplexed on a single lane in an Illumina HiSeq 2500.
GEO
GSM3874638
GEO Accession
GSM3874638