GSM8242955: LC_AD04; Homo sapiens; RNA-Seq
3mm biopsies were taken from atopic dermatitis patients (n=4) and healthy controls (n=3), Epidermis and dermis were separated after ON incubation and processed using the Miltenyi Epidermis Dissociation Kit or the Miltenyi Whol Skin Dissociation Kit, respectively. Single cell suspensions were stained and live LCs and T cells were sorted into 384-well plates containing vapor-lock (Qiagen) with 100-200 nl of primers, dNTPs and synthetic mRNA SpikeIns and were immediately centrifuged (2,000 x g) and frozen at -80⁰C Illumina sequencing libraries were prepared with the TruSeq small RNA primers (Illumina) and sequenced single end at 75-bp read length with 75,000 reads per cell on a NextSeq500 platform (Illumina). Sequencing reads were mapped against the reference human genome (GRCh38) using Burrows-Wheeler Alignment tool to create read counts