GSM9787406: Epidermal keratinocytes, Naked mole-rats Sample 3; Heterocephalus glaber; RNA-Seq
For RNA-Seq analysis of epidermal keratinocytes, NMR and mouse skin was digested overnight with Dispase II (5mg/ml) and separated epidermis was then digested with TrypLE Express. Detached keratinocytes were washed, labelled with anti-CD49f antibody and FACS-sorted. Keratinocytes were lysed in TRI Reagent and RNA was isolated using Direct-Zol Microprep Kit. Libraries were prepared using Next Generation Sequencing Library Preparation kit from NEBNext® according to manufacturer protocol. Ribosomal RNAs (rRNAs) were depleted using NEBNext® rRNA Depletion Kit v2 and libraries were prepared using NEBNext® Ultra™ II Directional RNA Library Prep Kit for Illumina®. After purifications, quality of the libraries was assessed on 5200 Fragment Analyzer with DNA Chip. For RNA-sequencing library preparation, concentration, quality and integrity of RNA was checked, and RNA with RIN>7 was used for the libraries.