GSM8767959: I30D; Mus musculus; RNA-Seq
Upon the euthanasia of mice, the mice were perfused with normal saline and tendons were dissected and placed in Dulbecco's modified Eagle's medium (low glucose; Thermo Fisher Scientific, USA) with 10% (vol/vol) fetal bovine serum (FBS, Gibco, USA) and 1% (vol/vol) penicillin-streptomycin (Thermo Fisher Scientific, USA). The specimens were rinsed twice with phosphate buffered saline (PBS) and minced, then subjected to pre-warmed dissociation solution (3 mg/mL Collagenase I (Gibco, USA) and 4 mg/mL Dispase II (Roche, Swiss) at 37 °C, 5%CO2 cell incubator for 2 hours. Every 30 minutes the tissues were gently blown by pipette. After digestion, 40-micron sterile strainers (Biosharp, China) were used to filter residue and the samples were centrifuged at 1,200 rpm for 5 min. To obtain single cell suspension, supernatant was discarded and 0.5mL PBS were added to resuspend the sediment. Single-cell suspensions (1×10^5 cells/mL) with PBS were loaded onto microwell chip using the Singleron Matrix® Single Cell Processing System. Barcoding beads collection, mRNA transcription, PCR amplification, fragmentation, ligation and library construction were conduct as per manufacture's protocol (GEXSCOPE® Single Cell RNA Library Kits, Singleron, China).
GEO
GSM8767959_r1