Single cell RNA-seq of skin biopsies from patients with cutaneous T cell lymphoma (CTCL)
Single cell RNA-seq of skin biopsies from patients with cutaneous T cell lymphoma (CTCL)
Samples were collected as biopsies from patients who have CTCL. Each biopsy was then digested into single cells by firstly separating the epidermis and dermis using dispase and then incubating each section with collagenase. Cells were FACS sorted before loading onto the 10x Chromium. During droplet encapsulation the cells are lyse. cDNA is made from the RNA of each cell by reverse transcription and then purified using Dyna beads. Libraries are constructed as per the 10x Genomics protocol: \"Enzymatic fragmentation and size selection are used to optimize the cDNA amplicon size prior to 5â€™ Gene Expression library construction. P5, P7, a sample index, and Illumina R2 sequence (read 2 primer sequence) are added via End Repair, A-tailing, Adaptor Ligation, and Sample Index PCR. The final libraries contain the P5 and P7 priming sites used in Illumina sequencers.\"