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A Complete Workflow from Single Cell Isolation to mRNA Sequencing Analysis

When performed on bulk samples, gene expression studies might yield only a limited understanding of complex samples. However, gene expression analyses performed on single cells can offer a powerful method to resolve sample heteroge¬neity and reveal hidden biology. BD’s fluorescence activated cell sorting (BD FACS™) technology can accurately identify and isolate single cells or a small number of cells for use in next generation sequencing (NGS) or transcriptome analyses. BD™ Precise assays enable examination of mRNA sequences for a large number of cells simultaneously at single-cell resolution. In this report, we describe a complete workflow from BD FACS identification and isolation of single cells to BD Precise sequencing library generation, sequencing, and data analysis. The single cells can be isolated using various BD FACS sorting instruments, from our entry-level sorters to our high-end sorters, BD FACSMelody,™ BD FACSAria™ III, BD FACSAria™ Fusion, BD SORP FACSAria™ Fusion, and BD Influx™. The index sorting feature of these sorters connects cells’ phenotypes to their molecular gene expression profiles. This paper shows a streamlined, fast method using a BD FACS sorter to select and isolate single cells for high-resolution NGS analysis. The combined workflow provides a power¬ful tool for studying complex biological samples at the resolution of individual cells.