GSM8386694: OV, WT, replicate 2, scRNAseq; Mus musculus; RNA-Seq
E10.5 OVs were pooled and digested in TryplE (Invitrogen) with 0.05% collagenase (Worthington Biochemical) and dispase (Worthington Biochemical) at 37°C for 30 min. 5U/ml DNaseI (Invitrogen) was added to the digestion mix after 15 min digestion. DPBS with 10% FBS was added to quench the digestion. Dissociated cells were spined at 300 g x 5 min, washed with DPBS with 0.04% BSA, and filtered with 40 μm strainer. Filtered cells were spined at 300 g x 5 min, washed with DPBS with 0.04% BSA, re-spined, and resuspended in 50 μl DPBS with 0.04% BSA for downstream processing. Single cell encapsulation and cDNA libraries were prepared with Chromium Next GEM Single Cell 3ʹ Reagent Kit v3.1 and Chromium Next GEM Chip G Single Cell Kit. Sequencing was carried out with Illumina NovaSeq 6000 for Pair-End 151bp sequencing.