GSM8462802: actl6a morphant, Smarca4a, FitCUT&RUN, rep3; Danio rerio; OTHER
To prepare samples for use with the FitCUT&RUN approach, the embryos at 30hpf were collected according to time after co-injection. For isolation of whole embryos cells, five 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 lysis solution and triturated gently five times with a 1000 μl pipette tip every 10 minutes until the end of 30 minutes of incubation. Finaly,The larvae were dissociated into single cells and then put in stop solution. Approximately 200000 viable cells per sample was resuspended for library construction. For FitCUT&RUN, the antibody incubation step was skipped, and the libraries were amplified from purified DNA using a Hyper Prep kits (KAPA Biosystems KK8504, USA). Finally, the libraries were sequenced by the Illumina Novaseq. FitCUT&RUN