Low Input RNA Sequencing of Freshly Isolated Muscle Stem Cells

Bulk RNA sequencing allows for the detection and quantification of mRNA transcripts from pooled cell populations, tissue sections, and biopsies. However, working with rare populations with limited amount of input requires specialized handling to ensure high-quality data. Here, we describe a detailed, optimized low-input protocol for bulk RNA sequencing on FACS-isolated satellite cells from mouse hindlimb skeletal muscle tissues. This protocol covers sample preparation, library preparation, sequencing, and data analysis.