GSM9195321: Adult_Tumor_Input_CUT&RUN_rep1; Danio rerio; OTHER
Embryos at 24hpf or tumors positive for GFP-2A-EF1 were dissected, cut into multiple pieces, and placed in pre-warmed Accumax at 30-32°C (Millipore Sigma, A7089), followed by mechanical dissociation by pipetting every 10 min for 1-1.5 hr. Cells were pelleted (200 × g, 10 min, 4 °C), and supernatant was removed and replaced with 1 mL of cold Hank's Buffer solution (0.25% BSA, 10mM HEPES, 1x HBBS). Cells were resuspended by pipetting up and down and pelleted again (376 × g, 5 min, 4 °C), and supernatant was removed and replaced with 1ml of 1x PBS. Pellets were resuspended and filtered through 40 um strainers (Corning, 431750). Cell number was assessed by automated cell counter. Cells were fixed in 1%PFA for 2 min. 100,000-200,000 cells per sample were aliquoted and pelleted for processing using CUT&RUN Assay Kit #86652 according to the manufacturer's protocol. Smaples were proccessed by Novogene. The DNA fragments were repaired, A-tailed and further ligated with Illumina adapter. The final DNA library was obtained by size selection and PCR amplification.