GSM7775319: EXOSC2 AID ESCs, RNA, sucrose F7, 0 hr; Mus musculus; RNA-Seq
Native nuclear lysates were fractioned by sucrose gradient centrifugation (6%-40% w/v). Add an equal amount of RNA spike-in (4 ERCC transcripts) to each fraction for normalization to cell number. 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. RNA-seq libraries were constructed by NEBNext Ultra II directional RNA library prep kits (NEB) following manufacturer's protocols.