title: GSM9328231: Mouse growth plate, rep2; Mus musculus; RNA-Seq
library_construction_protocol: The obtained slices were dissected from bone under a stereo microscope, ground into small fragments, transferred to fresh, ice-cold HBSS, and thoroughly pipetted to remove residual blood. Then HBSS was replaced with a digestion solution comprising Collagenase P (3 U/mL, Roche) and TrypLE™ Select Enzyme (1X, Thermo Fisher Scientific, Catalog No: A1217701) in HBSS. The tubes were incubated for 30–40 minutes at 37°C with shaking at 120 rpm. During digestion, the tissue fragments were disrupted every 10 minutes using a 1-mL pipette. The digestion was terminated by the addition of 10 mL of 2% FBS in cold HBSS. The mixture was centrifuged at 300 rpm for 10 minutes at 4°C. The supernatant was carefully removed, and the pellet was resuspended in 100 µL of 2% FBS in 1X PBS. Ice-cold methanol (400 µL) was added dropwise to the suspension, and the sample was placed on ice for 10 minutes. The cells were stored at -80°C for a maximum of six months prior to library preparation. 10X protocol