GSM9238024: A549 cells,DDX17-KD; Homo sapiens; ChIP-Seq
After the cells are digested, add serum-containing culture medium to stop digestion, and perform cell counting. For each sample, take 1-1.5x10^6 cells and centrifuge at 110xg for 5 minutes, discard the culture medium, wash once with 1xPBS, and then resuspend. According to the kit instructions, use lectin-coated magnetic beads to capture the cells, permeabilize the cells, and then add secondary antibodies corresponding to the primary antibodies (without HRP). Use Mg2* to activate Protein A/G-Tn5 transposase to cut the target fragment, terminate the reaction with proteinase K, and add adapters for amplification at both ends before purification and sequencing. Based on the analysis of the sequencing results, decide on the subsequent experiments. Using the Novogene library amplification kit, first prepare the components required for library amplification in a sterilized PCR tube: the components and their corresponding volumes are: 15 μl of resuspended DNA Extract Beads Pro, 25 μl of 2×CAM, 5 μl of N5XX, 5 μl of N7XX, totaling 50 μl. Then, gently pipette the mixed components to ensure they are well combined, and perform the reaction in the PCR machine according to the following program: 72°C for 3 minutes; 95°C for 3 minutes; then 98°C for 10 seconds, followed by 60°C for 5 seconds for 9 to 20 cycles; then 72°C for 1 minute, and hold at 4°C.