GSM9360606: GM05386_TGFb_rep3; Homo sapiens; RNA-Seq
RNA extraction was performed using a hybrid trizol-column protocol. Briefly, the upper aqueous phase after chloroform phase separation was then transferred and diluted with an equal volume of 70% ethanol, followed by transfer to a RNeasy mini spin column (Qiagen, 74104) as the manufacturer`s recommendation. RNA extracts were DNAse treated using the RNAse free DNAse set (Qiagen, 79254), and their quality checked by Agilent Bioanalyzer for the RNA integrity (RIN value). Library preparation and sequencing was performed by Genome Québec Libraries were generated from 20 ng of total RNA as following: mRNA enrichment and cDNA synthesis was achieved with theNEBNext Single Cell Low Input RNA Library Prep Kit for Illumina (New England BioLabs). Adapters and PCR primers were purchased from New England BioLabs. Libraries were quantified using the KAPA Library Quantification Kits - Complete kit (Universal) (Kapa Biosystems). Average size fragment was determined using a LabChip GX (PerkinElmer) instrument.