{"database":"GEO","file_versions":[{"headers":{"Content-Type":["application/json"]},"body":{"files":{"Other":["ftp://ftp.ncbi.nlm.nih.gov/geo/series/GSE347nnn/GSE347182/"]},"type":"primary"},"statusCodeValue":200,"statusCode":"OK"}],"scores":null,"additional":{"omics_type":["Transcriptomics"],"species":["Mus musculus"],"gds_type":["Expression profiling by high throughput sequencing"],"full_dataset_link":["https://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE347182"],"repository":["GEO"],"entry_type":["GSE"],"additional_accession":[]},"is_claimable":false,"name":"Gradient-Free Isolation of Murine Pancreatic Islets for Single-Cell RNA Sequencing","description":"Pancreatic islet isolation for single-cell RNA sequencing (scRNA-seq) requires extensive enzymatic and mechanical processing that may affect cell recovery and transcriptional state. We evaluated a practical gradient-free workflow for murine pancreatic islets from diabetic BKS.Cg-Dock7^m +/+ Lepr^db/J mice. The protocol combined intraductal collagenase perfusion, controlled digestion, stereomicroscope-guided manual islet selection without density-gradient purification, and gentle Accutase/EDTA dissociation. Compared with Ficoll-based purification, the gradient-free workflow omitted density-gradient preparation, gradient centrifugation, additional wash centrifugations, and associated transfer steps while yielding viable cells suitable for scRNA-seq. The resulting library met the applied QC criteria and retained the major endocrine populations. Stress-associated transcriptional differences between the two pooled libraries were descriptive only. Overall, the gradient-free workflow represents a feasible alternative to Ficoll-based purification for preparing murine pancreatic islets for scRNA-seq.","dates":{"publication":"2026/09/30"},"accession":"GSE347182","cross_references":{"GSM":["GSM10047090","GSM10047091"],"GPL":["24247"],"GSE":["347182"],"taxon":["Mus musculus"],"PMID":["[42776689]"]}}