GSM6383745: Chordoma tumor, CH8, miRNA; Homo sapiens; miRNA-Seq
Snap-frozen chordoma tissue samples were pulverized while frozen, the tissue powder was lysed immediately with TRIzolate Reagent (Sigma-Aldrich) (50 mg sample per 500 uL reagent), and the samples were homogenized with a rotor-stator homogenizer. An equal volume of chloroform was added to the homogenizates and vortexed, and the aqueous phase was separated using Phase Lock Gel™ (QuantaBio) (PLG 15 ml Heavy) tubes. RNA was isolated from the aqueous phase with the Direct-Zol RNA Miniprep kit (Zymo Research), according to the manufacturer's instructions, without DNase I treatment. RNA was prepared from the NP cell cultures with TRIzolate Reagent and Direct-Zol RNA Miniprep kit, according to the manufacturer's instructions. RNA quality was checked with Nanodrop 1000 spectrophotometer and Agilent 2100 Bioanalyzer. Sequencing libraries were prepared with the Illumina TruSeq Small RNA SamplePrep kit. miRNA-seq, 1x 50 bp