GSM9068158: Tooth germ, gene modification, Edarfl/fl, rep1; Mus musculus; RNA-Seq
The tissue samples were processed as described below. Briefly, samples were first washed with 1640 culture solution, minced into small pieces (approximately 1mm3) on ice and enzymatically digested with 1mg/ml Collagenase I , 0.5mg/ml Collagenase II, 0.4% 1mg/ml Dispase, 50U/mL DNase I for 30~40 min at 37°C, the supernatant was added to an equal volume of 1640 culture solution containing 10% FBS to terminate digestion; After digestion, samples were sieved through a 70µm cell strainer. After the supernatant was removed, the pelleted cells were suspended in red blood cell lysis buffer (Miltenyi Biotec) to lyse red blood cells. Dissociated single cells were then stained with AO/PI for viability assessment using Countstar Fluorescence Cell Analyzer. The single-cell suspension was further enriched with a MACS dead cell removal kit (Miltenyi Biotec). BD Rhapsody system  were used to capture the single cells and construct the scRNA-Seq libraries