GSM5216961: RNA_seq_basal_cells_WT_cont_ex2 [Cont_2]; Mus musculus; RNA-Seq
Subcutaneous fat was removed from the skin with a scalpel, and the whole skin was trypsinized at 37 °C for an hour to separate the epidermis and the dermis. The cell suspensions were filtered through strainers (70-μm, BD Falcon), collected by centrifugation (300 × g, 5 min) and suspended in Dulbecco's modified Eagle's medium (Ca-). Cell suspensions were incubated with Pacific Blue anti-human/mouse CD49f (Biolegend) antibody and eFluor660 anti-mouse CD34 (Biolegend) antibody for 30 min on ice, washed twice with PBS and subjected to flow cytometric analysis. Cell isolation was performed on a FACSAria Ⅲ sorter. RNA was extracted from CD49f positive and CD34 negative cells using the RNeasy Micro Kit (Qiagen). Quality RNA samples were selected (RIN>7). RNA-seq libraries were generated using the TruSeq Stranded mRNA library prep kit (Illumina) and the libraries were sequenced on NextSeq 500 (Illumina).