GSM9598479: Single cells from islets, WT2 GlcN; Mus musculus; RNA-Seq
Following treatment, pancreatic islets were dissociated into single-cell suspension using Accutase. Nuclei were subsequently isolated according to the 10x Genomics Demonstrated Protocol ("Nuclei Isolation for Single Cell Multiome ATAC + Gene Expression Sequencing”, CG000365"). During the lysis buffer incubation, manual dissociation was performed by passing the suspension through a 30G needle with a 1-mL syringe to facilitate nuclear release and isolation The scRNA libraries were prepared using the Chromium Next GEM Single Cell Multiome ATAC and Gene expression reagent Kit (10x Genomics). Nuclei were diluted in 5 ul and proceed to the transposition mix and incubated for one hour at 37⁰C and according to the company manufacture (10x Genomics). Then the mix was separated into droplet emulsion using a Chromium X machine (10x Genomics) and libraries were prepare according to the manufacturer's protocol. The scRNA-seq libraries were sequenced on the Illumina Novaseq6000 sequencing system (Illumina) as a 28-bp and 90-bp paired-ended.