GSM8322482: sample pool 3, run HGYVNCCX2; Mus musculus; RNA-Seq
Single cells were liberated from mineralizing cultures via incubations with 60 mM ethylenediaminetetraacetic acid pH 7.4 (EDTA [Thermo Fisher Scientific], made in DPBS), 8 mg/mL collagenase (Gibco) in HBSS/4 mM CaCl2 (Fisher), and 0.25% trypsin–EDTA (Gibco). After single-cell isolation, cells from mice were pooled into groups containing cells from five mice total and concentrated to 800 cells/μL in PBS supplemented with 0.1% BSA (bovine serum albumin). Pooled single cells were prepared for sequencing using the 10× Chromium Controller (10× Genomics, Pleasanton, CA, USA) with the Single Cell 3' v2 reagent kit, according to the manufacturer's protocol. Libraries were sequenced on the NextSeq500 (Illumina, San Diego, CA, USA).