GSM9019599: E6.5 ex utero cultured for 2 days mouse embryos in hypoxic condition (2-5% O2); Mus musculus; RNA-Seq
The embryos were pooled according to the treatment conditions and dissociated in TrypLE at 37 degrees for 10 minutes with pipetting every 5 minutes. The dissociated cells were mixed with recovery medium (10% FBS in low glucose DMEM) and spun down for 5 minutes at 40 rcf, followed by one time wash in PBS, and ultimately placed in PBS with DRAQ5 and DAPI, subject to On-chip sort microfluidic chip sorter (On-chip Biotechnologies) to enrich for viable cells (DRAQ5+DAPI-). The resulting sorted suspensions in 1X PBS and 1% BSA were put into separate lanes for GEM generation to the 10X Chromium GEM-X Single Cell v4 3' protocol Samples were processed according to 10X Chromium GEM-X 3' v4 protocol