GSM6533094: scD1, wt replicate 2, scRNAseq; Mus musculus; RNA-Seq
Embryos were harvested at E8.0 in ice-cold phosphate buffered saline (PBS) containing 10% (v/v) of fetal bovine serum (FBS/PBS). Embryos aged between 0-1 somite were selected for further analysis and the extraembryonic tissue was removed. Embryos were washed with Ca+2/Mg+2- free Dulbecco's phosphate buffered saline DPBS. Embryos were incubated in TrypLETM Express (Gibco) at 37oC for 6-7 min with intermittent pipetting using wide bore tips. Once embryos were completely dissociated, the enzyme was quenched using 10% FBS/DPBS (Ca+2/Mg2+ free) before the cells were resuspended in Ca2+/Mg2+ free DPBS containing 5% (v/v) endotoxin free FBS. Cell viability (i.e., lack of dead cells), number and size were confirmed with a hemocytometer. Library was performed according to the manufacter's instructions (single cell 3' library v3 protocol, 10x Genomics).