Instrument: Illumina NovaSeq 6000
Strategy: RNA-Seq
Source: TRANSCRIPTOMIC
Selection: cDNA
Layout: PAIRED
Construction protocol: Embryos were harvested from the uterus and dissected in cold dissecting buffer (Dulbecco's phosphate buffered saline with 10% fetal bovine serum and antibiotics). Dissected tissues were dissociated in 0.125% collagenase type I (Sigma-Aldrich) and 1 U Dispase (Stemcell Technologies) at 37°C for 30 min. The samples were reduced to single cell suspension by tripsin digestion, washed with cold PBS, and passed through 40mm cell strainer. Embryonic cells were lysed and single cell libraries prepared by following protocols provided by 10x Genomics. For the ESC differentiation experiment, we utilized the MULTI-seq sample barcoding and library preparation protocol from the McGinnis lab (McGinnis et al. 2019).