title: GSM3536485: 3'mRNAseq_wt_rep1; Homo sapiens; RNA-Seq
library_construction_protocol: Nuclei were prepared by incubation of cells 5 mins on ice in HLB (10 mM Tris pH 7.5, 10 mM NaCl, 2.5 mM MgCl2, 0.5% NP40), underlayering with HLB+NS buffer (10 mM Tris pH 7.5, 10 mM NaCl, 2.5 mM MgCl2, 0.5% NP40, 10% sucrose) and centrifugation 420 g for 5 min at 4oC. RNA was purified from the nuclei using TRI reagent (Sigma) according to manufacturer's instructions. Residual DNA was digested using 4U Turbo DNase (Life Tech) for 10 min at 37oC followed by proteinase K digestion for 10 min at 37oC. TRI reagent purification and DNase digestion were repeated. RNA was further acid phenol/chloroform and chloroform extracted, followed by ethanol precipitation. The purified RNA was then resuspended in ultrapure water. Libraries were prepared according to the protocols of QuantSeq 3'mRNA-Seq library prep kit REV for Illumina (LEXOGEN Cat# SKU 016.24)
attributes.tag: GEO Accession
attributes.value: GSM3536485