title: GSM8190289: R141, T1D, rep 1; Homo sapiens; RNA-Seq
library_construction_protocol: Pancreatic islets and islet-depleted tissue were dispersed into single-cells by dynamic incubation with 0.025% trypsin (Gibco) supplemented with 10 mg/mL DNase (Pulmozyme, Genentech) for 6–7 min and filtered through a 40 μm cell-strainer to remove undigested material. For automated CEL-seq2, cells were FACS sorted into 384-well cell capture plates. Cells were lysed by heat (65 Celsius degrees) followed by a second-strand cDNA synthesis. For each plate, all barcoded material was amplified by in vitro transcription. Cel-Seq2 workflow was followed to generate the cDNA library using TruSeq small RNA primers (Illumina). The DNA library was paired-end sequenced using Illumina Nextseq 500, high output, with a 1x75 bp Illumina kit (read 1: 26 cycles, index read: 6 cycles, read 2: 60 cycles). For 10X, cells were loaded, and the sequencing libraries were prepared following the standard 10x Genomics 3' V3 chemistry protocol