GSM7804816: EXOSC2 AID ESCs, RNA, chrRNP, rep1, 0 hr; Mus musculus; RNA-Seq
Cells were fractionated by sequential lysis and centrifugation. We first isolated the cytosolic fraction ('cyto') by 0.15% NP-40, and then used salt and detergents (0.15 M NaCl, 0.5 M urea and 0.5% NP40) to partition nuclear constituents. The soluble nucleoplasmic fraction ('np') with greater biochemical extraction captured the highly dynamic RNP mesh components, whereas the insoluble fraction contained chromatin and less extractable RNP mesh components, designated as the 'chr/RNPmesh'. Add the RNA spike-in (a combination of GFP, mCherry, lacZ, and Luciferase transcripts) directly to the samples of each subcellular fraction according to number of cells for normalization. Total RNA were extracted by TRIzol reagent according to manufacturer's instruction. Purified total RNA were subjected to ribosomal RNA depletion by Ribo-Zero rRNA removal kits (Illumina) according to manufacturer's instruction.