GSM4212554: SS-nano; Homo sapiens; RNA-Seq
Total RNA was isolated from a 6-well by directly lysing the cells with TRIzol Reagent according to the manufacturer's protocol (ThermoFisher) and then further purified using RNeasy columns (Qiagen, Germantown, MD, USA). The presence of contamination with chemicals was assessed by Nanodrop using 260/280 and 260/230 ratios. Quantification of total RNA was made by QuBit (Applied Biosystems, Waltham, MA, USA) and 1000 ng of total RNA was used for library preparation. Quality of total RNA was assessed with the BioAnalyzer Nano (Agilent, Santa Clara, CA, USA) and all samples had a RIN above 9. Library preparation was done with the KAPA mRNAseq stranded kit (KAPA, Cat no. KK8420). Ligation was made with 11 nM final concentration of Illumina index and 9 PCR cycles were required to amplify cDNA libraries. Libraries were quantified by QuBit and BioAnalyzer. All libraries were diluted to 10 nM and normalized by qPCR using the KAPA library quantification kit (KAPA; Cat no. KK4973). Libraries were pooled to equimolar concentration.
GEO Accession
GSM4212554