GSM2869255: ActD_timecourse_rep2_time240; Mus musculus; RNA-Seq
300,000 mESCs were seeded per well of 2 6-well dishes and grown for 48 hr in serum + LIF medium. The medium was replaced by fresh medium containing 5 μM Actinomycin D (from a 5 mg/ml stock in DMSO) and the cells incubated for 30, 90 or 240 min. A mock treatment (240 min = "time 0" samples) was included, using medium with the same amount of DMSO but no Actinomycin D. After the indicated times, the wells were washed twice with 37 °C PBS, and RNA extracted using the Agilent Absolutely RNA miniprep kit. ERCC RNA spike-ins were added to the lysis buffer (1.7 μl of a 1:10 dilution per sample) before it was added to the cells. Ribosomal RNA was then depleted with the ribo-zero rRNA removal kit and high throughput sequencing libraries prepared using the ScriptSeq V2 RNA-Seq Library Preparation Kit.
GEO Accession
GSM2869255