GSM9752993: WT FAPs isolated from ND muscle; Mus musculus; RNA-Seq
Ten TA muscles from five male mice per strain were minced in wash medium (F-10 with 10% FBS and 1% penicillin–streptomycin) and digested for 1 h at 37 °C in wash medium containing Collagenase II, Dispase II, and 1% BSA. The suspension was filtered twice through 40 µm strainers, treated with RBC lysis buffer for 3 min, and stained with Propidium Iodide (Invitrogen) to exclude dead cells. Single-cell libraries were constructed using the Chromium GEM-X Single Cell 3′ Reagent Kit v4 according to the manufacturer's instructions. Reverse transcription was performed within the GEMs to generate barcoded, full-length cDNA. Following emulsion breakage, cDNA was purified and amplified. Final sequencing libraries were prepared via cDNA fragmentation, end-repair, A-tailing, and ligation of Illumina-compatible dual-index adapters. Libraries were multiplexed and sequenced on an Illumina NovaSeq X platform (UF ICBR NextGen DNA Sequencing Core) to achieve an average sequencing depth of approximately 40,000 reads per cell.