title: GSM4768936: Amino acid suplementation_rep1; Homo sapiens; ncRNA-Seq
library_construction_protocol: RNA was extracted once the cells reached 70-90% confluency. Small RNA was isolated using Isogen2. After ethanol precipitation, periodate oxidation of small RNA was carried out in 100 mM NaOAc/HOAc, pH 5.2, freshly prepared 50 mM NaIO4 at 27°C for 30 minutes. The reaction was quenched with 100 mM glucose for 5 minutes at 27°C. tRNA was then purified by denaturing 15% PAGE(5M Urea, 1× TBE), and eluted from the gel overnight in TE.   For β-elimination and deacylation, tRNA was treated in 150 mM Tris-HCl, pH 9.0, at 37 °C for 30 min. tRNA was demethylated as previously described (Zheng 2015). RNA libraries were prepared for sequencing using NEBNext Small RNA Library Prep Set for Illumina.
attributes.tag: GEO Accession
attributes.value: GSM4768936