title: GSM7324963: CUTTag_antiBRD9_Gastricpremalignant organoids, Trp53-/-;sgPten;sgArid1a_rep2; Mus musculus; OTHER
library_construction_protocol: Organoids (3-10X104 cells/each group) were harvested in 1.5ml tubes for CUT&Tag assay using NOovoNGS CUT&Tag 3.0 High-Sensitivity Kit (Novoprotein, Cat. N259-YH01-01A). 10ul ConA beads were added into tube and incubated with cells for 10min at 25°C. The supernatant was removed and added primary antibody buffer with proper primary antibody for 2h at 25°C or overnight at 4°C. The supernatant was removed and added secondary antibody buffer for 1h at 25°C. Beads were washed for 2 times. The chromatin was combined with ChiTag TM2.0 Transposome, then fragmented using Tagmentation Buffer for 1h at 37°C. ChIP DNA was purified using DNA Clean Beads and amplified using the PCR system from this kit. PCR conditions: 72 °C for 3 min, 98 °C for 30 s, followed by 10-25 cycles of 98 °C for 15 s, 60°C for 15 s and 72 °C for 10 s, 72 °C for 2min. Amplified libraries were purified with the DNA Clean Beads.. The purified ChIP DNA was used for library preparation.