GSM8147003: Evf2++ rep1 scRNA-seq; Mus musculus; RNA-Seq
For scRNAseq, E13.5GE dissections from Evf2+/+ and Evf2TS/TS embryos were collected into ice cold Leibovitz's L-15 medium and 1% fetal bovine serum. For the single cell dissociation, papain dissociation kit, protocol II (Worthington Biochemical, LK003150). In the last step, the supernatant was removed and pelleted cells were resuspended in 1ml pre-chilled L15 medium + 1% fetal bovine serum.  Cells were counted using a Luna cell counter, and viability determined to be ~90%. The 10X Genomics microfluidics system and the 10X Genomics Chromium Single Cell 3' Reagent Kit v3 were used for single cell separation and library preparation, following all protocol recommendations, including PCR cycles (12 cycles for cDNA amplification of 5000 cells and 14 cycles for index PCR.