GSM6043087: Batch 1 WT; Mus musculus; RNA-Seq
E8.5 embryos were pooled and washed twice in chilled PBS before adding 100µl 1X Trypsin/EDTA and incubating at 37ºC for 10 minutes. Pools were then pipetted up and down gently 20 times using a P200 pipet to create a single-cell suspension and the trypsin was inactivated by addition of 900µl chilled L-15 medium + 2% FBS. Suspensions were then passed through a 50µm filter to remove any cell clumps/larger debris into 5ml round-bottomed polypropylene tubes and stained with viability dye. Live cells were sorted using a BD Melody flow sorter into a 1.5ml microcentrifuge tube containing L-15 medium + 2% FBS to a final volume of 47µl ready for single-cell sequencing Library was performed according to the manufacter's instructions (single cell 3' v3 protocol, 10x Genomics).