GSM8462821: pronephric duct, ctrl morpholino injected, 30hpf, rep1; Danio rerio; ATAC-seq
For isolation of pronephros ducts, 30 hpf. Fifteen zebrafish embryos were incubated in 10 mmol/ml DTT in egg water for 1 hour at room temperature. They were then incubated at 37 °C in 5 mg/ml collagenase I in DMEM/F12 and triturated gently five times with a 1000 μl pipette tip every 10 minutes until the end of 30 minutes of incubation. The larvae were then put in DMEM/F12, so that the disintegrated, approximately 10 pronephric duct fragments were collected by visual identification according to the GFP fluorescence and dissociated into single cells. 10000-20000 viable cells per sample was resuspended for library construction. The Hyperactive ATAC-Seq Library Prep Kit for Illumina (Vazyme, China ) was used to prepare an ATAC-seq library following the manufacturer's suggested procedures.