title: Mouse_RA_treatment_15dpi_ear_pinna_scRNA-seq_10x
design: The ear hole tissues were collected using biopsy punches and washed with ice-cold PBS. Subsequently, the tissues were dissected into small pieces quickly and transferred into tubes with Collagenase dissociation buffer containing 2mg/ml Collagenase type I (Sigma, C2674) and 100ug/ml DNase I (Roche, 10104159001). After incubating at 37 , the dissociated cell suspension was filtered through a 70 m filter. The remaining incompletely digested tissue pellets were further dissociated with TrypLE (Gibco, 12605010) and then filtered and mixed with dissociated cells digested with collagenase. For separate living cells, the dissociated cell mixtures were resuspended with ice-cold PBS containing 5% fetal bovine serum (FBS) and incubated with Propidium Iodide (PI) to stain dead cells. The PI-negative cells were sorted using BD FACSAria II Flow cytometry. To ensure single-cell sample quality, the proportion of living cells was evaluated by Trypan blue staining before library preparation with the 10X Genomics platform.【NOTE】This raw data is too large to download using prefetch with default parameters. Please using this command : 'prefetch --type any SRR329xxxxx --max-size 60000000' to download them. The SRA run is not empty!!!!!!!! Files over 100G have been splited into two files(part1, part2) for uploading. Please merge them first before running downstream pipelines.