GSM6939645: Prom1_BcatGOF; Mus musculus; RNA-Seq
Whole bile ducts were isolated and micro dissected in petri dishes to remove all adjoining tissue. Ducts were placed in 1 ml of HBSS with 1.8 mM Ca, 0.8 mM Mg, 1 mg dispase (Roche), 2 mg protease from Bacillus licheniformis (Sigma) and cut into small pieces with a microdissecting scissors. These were incubated for 15 minutes at 37 C. This solution was removed and the remaining bile duct chunks were placed in a solution with 1x PBS, 5mM EDTA, and 0.5% BSA and incubated at 37 C for another 10 minutes. This was filtered through a 100 μm filter. This single cell suspension was washed twice with a 1x PBS, 0.04% BSA solution prior to GEM and library generation according to manufacturer's protocol (10X Genomics, Chromium Single Cell 3' Reagent Kits User Guide, v3 Chemistry)