GSM4120721: ATAC HH6 rep1; Gallus gallus; ATAC-seq
For isolation of neural crest cells, embryos were transfected with the enhancer reporter construct PTK-AP2aE1-eGPF. To obtain neural crest cells from different stages, embryos were cultured until HH6 (8 hours) and HH9 (13-14hours), and screened for robust GFP expression in neural crest cells. Embryo heads were dissected and incubated in Accumax (Accutase, SCR006) cell dissociation solution, for 40 minutes at RT under mild agitation. At least 20,000 GFP+ cells from each stage were sorted into 200uL of HANKS buffer supplemented with 0.5% BSA. ATAC-seq library preparation was performed as previously described (Buenrostro et al., 2015). Neural plate border/neural crest dissections were performed on HH6/HH9 embryos (10 embryos/ 20 neural folds per experiment). Cells were dissociated in Accumax (Accutase, SCR006) cell dissociation solution for 20 minutes at RT under mild agitation. CUT&RUN experiments were carried out as described(Skene and Henikoff, 2017).
GEO Accession
GSM4120721