GSM9791370: Dermatomyositis lesional skin, DM1; Homo sapiens; RNA-Seq
Skin biopsies were collected by 4-mm punch biopsy after local anesthesia with 1% lidocaine with epinephrine, placed in formalin, and embedded in paraffin. For each sample, four FFPE curls (25 μm each) were dissociated following the FLEX protocol (CG000632 RevA, 10X Genomics) using the Miltenyi Biotec FFPE Tissue Dissociation Kit. Cells from each sample were hybridized with a unique Probe Barcode according to the "Chromium Fixed RNA Profiling Reagent Kits for Multiplexed Samples" user guide (CG000527, 10X Genomics). Post-hybridization, cells from 16 samples were washed, counted, and pooled in equal proportions. Approximately 200,000 cells from the pooled suspension were loaded onto a Chromium Q chip (PN-1000422, 10X Genomics). Sequencing libraries were prepared and sequenced on an Illumina NovaSeq platform using paired-end dual-indexing (28 cycles for Read 1, 10 cycles for i7, 10 cycles for i5, and 90 cycles for Read 2).