GSM6523199: Littermate control uninjured [Day_7_WT_uninjured_S4]; Mus musculus; RNA-Seq
Tenotomy injury site, which includes the Achilles' tendon, from the tendon-muscular junction at the gastrocnemius muscle to the calcaneal enthesis, and surrounding tissue posterior to the tibia, was dissected one week following injury. Tissue was finely minced then digested for 20 minutes in 750 ul/ml type 1 collagenase and 7 U/ml dispase II mixture (Gibco) with Roswell Park Memorial Institute (RPMI) medium at 37oC with agitation at 160 rpm. Digestions were quenched with 2% FBS and filtered through 40 micron strainers. The resulting cell suspension was washed in 2% FBS in PBS. Cells were counted and resuspended at 1000-12000 cells/ul. Viability was determined using Countess II (Thermo Fisher Scientific). Samples with greater than 85% viability were submitted for sequencing. 50K reads per cell. 3 Prime gene expression. 150 cycle high output (500 M). Read 1 Run Configuration: 28bp. Read 2 Run Configuration: 98bp. i7: 10bp. i5: 10bp.