title: GSM8377726: NTS, scRNA seq; Mus musculus; RNA-Seq
library_construction_protocol: Achilles's tendon enthesis of the SKG mice were dissected and transferred to sterile centrifuge tubes, the tissues were cut into 2 mm tissue blocks and digested with 0.5 % collagenase type II for 2 hours in a 37℃constant temperature shaker. The cell suspension was filtered through a 40 µm cell strainer, then centrifuged at 400×g for 6 min at room temperature and discard the supernatant to collect dissociated cells. Red blood cell lysis buffer was added to resuspend cells and incubated for 5min at room temperature. The cell suspension was centrifuged again at 400×g for 6 min at room temperature and the Dead Cell Removal Kit was used to remove dead cell. Cell viability was assessed by trypan blue, sample with greater than 80% cell viability were qualified to perform 10× Genomics single-cell RNA sequencing. Single-cell suspension was loaded to a Chromium Controller and partitioned into Gel Bead-In-Emulsions (GEMs) by the microfluidic cross-system according to the manufacturer's protocol. Reverse transcription reaction was performed to synthesize cDNA, followed by fragmentation, adapter ligation, PCR amplification and products purification. The libraries were sequenced on Illumina NovaSeq 6000