GSM9684753: Human islets, donor 3; Homo sapiens; RNA-Seq
Human islets (2,000 IEQ/donor) were dissociated into sin-gle cells using TrypLE Express containing 1 mM EDTA(ThermoFisher Scientiﬁc, Cat. No. 12605028) at 37_x000E_C for 12–15 min. Cells were washed twice with PBS containing 0.1%human serum albumin (Baxalta, NDC 0944-0493-01). Five toseven thousand cells from each donor preparation were usedfor droplet-based single-cell mRNA sequencing analysis onthe 10x Genomics Chromium platform of 30 Library and Gelbead Kit V3.1 Single-cell gene expression libraries were generated using the 10x Genomics Chromium Single Cell Gene Expression kit following the manufacturer's protocol. Single cells were encapsulated with barcoded gel beads in emulsion (GEMs), and mRNA molecules were captured and reverse transcribed using oligo-dT primers containing cell barcodes and UMIs. Barcoded cDNA was amplified by PCR, followed by fragmentation, adapter ligation, and index PCR to generate sequencing-ready libraries. Libraries were assessed for quality and sequenced on an Illumina sequencing platform.