GSM7927715: OVDMSO; Homo sapiens; RNA-Seq
Total RNA was isolated using Trizol reagent (Life technologies, USA) and 250  ng of total RNA was used for the construction of sequencing libraries Library was prepared using the NEBNext® Ultra™ II RNA Library Prep Kit for Illumina (Catalog: E7775S, New England Biolabs). mRNAs were primed with NEBNext Random Primers and chemically fragmented in a magnesium-based buffer in order to get an inserts of ~200 nucleotides. The fragmented mRNAs were reverse transcribed to form cDNA and the first strand cDNA reactions were converted to ds DNA. The double stranded cDNA fragments obtained were cleaned up by using 1.8X of AMPure XP beads (Catalog: A63881, Beckman Coulter). The library concentration was determined in a Qubit.3 Fluorometer (Catalog: Q33216, Life technologies using The Qubit dsDNA HS (High Sensitivity) Assay Kit (Catalog: Q32854, ThermoFisher Scientific). The library quality assessment was done using Agilent D5000 ScreenTape System (Catalog: 5067-5588, Agilent) and Agilent D1000 ScreenTape System (Catalog: 5067- 5582, Agilent) in a 4150 TapeStation System (Catalog: G2992AA, Agilent).