title: GSM8333043: mononuclear cells, quadriceps, day 3 post BaCl2-induced acute muscle injury; Mus musculus; RNA-Seq
library_construction_protocol: Cryo-preserved cells were used directely for library construction for scRNA-seq. Cells were thawed, washed, and resuspended at 1,000 cells/ul for processing 10x scRNAseq v3.1 protocol. 10X Genomics Single Cell 3' Reagent Kits V3.1, following the User Guidelines. Briefly, GEM RNAseq was performed as described (10x Genomics, Pleasanton, CA), following the Single Cell 3' Reagents Kits V3 User Guides. Cells were filtered, counted on a Nexcelome instrument, and the final concentration was set at 1,000 cells/μl for cells. The amount of cells or nuclei loaded on the chip was determined based on the 10X Genomics protocol. The 10X chip (Chromium Single Cell 3' v3 chip kit B PN-2000060) was loaded to target 5,000 cells final. Reverse-transcription was performed in the emulsion and cDNA was amplified before library construction following the Chromium protocols v3. Each library was tagged with a different index for multiplexing (Chromium i7 Multiplex kit PN-12062). Quality control and quantification of the amplified cDNA were assessed on a Bioanalyzer (High-Sensitivity DNA Bioanalyzer kit). Sequencing was carried out at the New York Genome Center on Illumina Novaseq using  98+26 paired-end reads.