GSM8774550: WT 36_2; Mus musculus; RNA-Seq
RNA from cells was extracted using commercial kits (MACHEREY-NAGEL GmbH & Co. KG) according to the provided instructions. High-sensitivity DNA chip (Agilent Bioanalyzer) were used to check library size distribution. All cDNA concentrations were quantified using the Quant-iT PicoGreen dsDNA Assay Kit (Thermo Scientific). Tagmentation was carried out in 2 µl, consisting of 1× tagmentation buffer (10 mM Tris, pH 7.5, 5 mM MgCl2, 5% DMF), 0.08–0.1 µl ATM (Illumina XT DNA sample preparation kit), 1 µl cDNA and water. Library amplification of the tagmented samples was performed using 1.5 µl custom designed Nextera index primers. After tagmentation, samples were pooled, and the pool was purified with Ampure XP beads. Libraries were sequenced at the 75-bp single end, 50-bp single end or 150-bp paired end on a high-output flow cell using an Illumina NextSeq500 instrument, an Illumina HiSeq3000 instrument or a NovaSeq S4 flow cell 150-bp paired end.