GSM8442441: miR-342 mice - pool 1; Mus musculus; RNA-Seq
Cells were sorted with the Flow cytometry (FC) before sequencing. Organs were smashed into small pieces and then incubated 15min at 37°C in agitation with a tissue digestion solution composed of 1 mL IMDM (Corning) supplemented with 0.35 mg/mL collagenase type IV (Sigma-Aldrich/ SCR103), 1 mg/ml dispase II (Gibco/ 17105041) and 0.2 mg/ml DNAse (Roche/ 11284932001). The tissue was then further dissociated by pipetting and filtered using 40 μm cell strainers (Corning/ 352340). Single cell solution was washed with 30 mL of autoMACS Running Buffer (Miltenyi Biotec/130-940 091-221) and pelleted for 5min 300g 4°C to remove excess enzymes. Samples were transferred to FACS tubes, stained with surface antibody and hashtags for sample identification. Then mOrange+ cells were sorted. Library was constructed using a microfluidics-based approach on Chromium Controller (10X Genomics) using the Chromium Next GEM Single Cell 5' Reagent Kits v2 (Dual Index) (10X Genomics) according to the manufacturer's instructions. For each mouse, 50,000 mOrange+ cells were sorted.  We then loaded 15-20,000 cells belonging to 3 mice from the same group onto the Chromium Controller (10X Genomics).