title: GSM5788503: P6_S13_NORMAL; Homo sapiens; RNA-Seq
library_construction_protocol: All fresh skin samples were gently washed in RPMI 1640 after removing crust, subcutaneous fat and necrotic tissue with surgical scissors and cutting the tissue into small pieces of 2–4 mm in a sterile tissue-culture dish. For tumor samples digestion was performed using tumor dissociation kit for human (130-095-929, MACS Miltenyi Biotec), mechanical dissociation using gentleMacs Dissociator running the gentleMACS program h_tumor_01. In order to capture enough keratinocytes in AK samples for subsequent research, we separate the epidermal tissue from the dermis and then dissociated to single-cell suspensions by combining mechanical dissociation with enzymatic degradation of the extracellular adhesion proteins using Epidermis Dissociation Kit for Human (130-103-464, MACS Miltenyi Biotec). The dissociated cell suspension was strained with a 40μm filter (BD Falcon), and treated with Red Blood Cell Lysis Solution (130-103-183, MACS Miltenyi Biotec) and dead cell removal using the Dead Cell Removal Kit (130-090-101, MACS Miltenyi Biotec) to confirm cell viability >85% with trypan blue staining (Invitrogen). All samples were processed as per manufacturer's instructions. Sorted cells were centrifuged and resuspended in PBS + 0.04% BSA (Gibco) to a final cell concentration of 700-1200 cells/μL as determined by hemacytometer. Chromium Single Cell 3' v3 (10X Genomics) library preparation kit according to the manufacturer's protocol