Optimizing Single-Cell Long-Read Sequencing for Enhanced Isoform Detection in Pancreatic Islets [bulk_islet]

The goal of this study was to optimize single-cell long-read RNA sequencing in pancreatic islets to enable accurate isoform-level gene expression analysis. We aimed to overcome key technical limitations—including insufficient read lengths, high error rates, and transcript abundance bias—that have hindered long-read sequencing in single cells. Specifically, we compared 5' and 3' library preparation strategies and evaluated transcript depletion methods to improve detection of low-abundance isoforms. Our objective was to establish a robust protocol that enhances isoform identification and reveals differential transcript usage across pancreatic islet cell types. Overall design: Bulk RNA-seq libraries og C57BL/6 mouse pancreatic islets, with and without insulin depletion. Two replicates of each.