GSM3993061: U1-3_cell; Mus musculus; ncRNA-Seq
Subcellular fractionation was performed as previously described with following modifications45. Cytoplasmic buffer containing 0.05% or 0.15% NP-40 was used for isolating cytoplasmic fraction of mESC or 293T cells respectively. The whole sample of each subcellular fraction was spiked and mixed thoroughly with 5 µl 0.1 ng/µl spike-in RNA [contain two in vitro transcribed RNA: lacZ and mCherry, For polyA RNA-seq or SLAM-seq, ERCC spike-in (Thermo Fisher) was used]. For total RNA-seq, ribosomal RNAs were removed through Ribo-Zero rRNA Removal Kits (Epicentre), for polyA RNA-seq, polyA RNAs were isolated through Dynabeads mRNA Purification Kit (Thermo Fisher). RNA-seq libraries were constructed through NEBNext Ultra II Directional RNA Library Prep Kit (NEB). Subcellular fractionation was performed as previously described with following modifications45. Cytoplasmic buffer containing 0.05% or 0.15% NP-40 was used for isolating cytoplasmic fraction of mESC or 293T cells respectively. The whole sample of each subcellular fraction was spiked and mixed thoroughly with 5 µl 0.1 ng/µl spike-in RNA [contain two in vitro transcribed RNA: lacZ and mCherry, For polyA RNA-seq or SLAM-seq, ERCC spike-in (Thermo Fisher) was used]. For total RNA-seq, ribosomal RNAs were removed through Ribo-Zero rRNA Removal Kits (Epicentre), for polyA RNA-seq, polyA RNAs were isolated through Dynabeads mRNA Purification Kit (Thermo Fisher). RNA-seq libraries were constructed through NEBNext Ultra II Directional RNA Library Prep Kit (NEB).
GEO Accession
GSM3993061