GSM9630894: Cytosolic RNA-seq mouse embryonic stem cell, 6 h time point, biological replicate 7; Mus musculus; RNA-Seq
Cells were dissociated with Accutase and pelleted by centrifugation. Cell lysates were separated into cytoplasmic and nuclear fractions using the SurePrep kit (Fisher Scientific) according to the manufacturer's instructions. RNA was immediately extracted from each fraction using the RNeasy kit (Qiagen) and stored at −80°C until sequencing. Paired-end strand-specific mRNA-seq libraries were prepared from cytoplasmic (cyR) and nuclear (nuR) RNA fractions using Illumina TruSeq Stranded mRNA Sample Preparation Kit according to the manufacturer's protocol. Libraries were sequenced on an Illumina HiSeq 4000 instrument with 150 bp paired-end reads and a read depth of approximately 10 million reads per sample per strand.