GSM6716953: LNCaP_METTL3Abcam; Homo sapiens; OTHER
100,000 cells and the HyperactiveTM In-Site ChIP Library Prep Kit (Vazyme Biotech, TD902) were used according to manufacturer's instructions. ChIP-seq libraries were prepared using the HyperactiveTM In-Site ChIP Library Prep Kit (Vazyme Biotech, TD902). Cells suspended in 100μL Wash Buffer were incubated with 10μL pre-washed ConA beads for 10 min at room temperature (RT). Primary antibody was added and incubated for 2h at RT. After washing with Dig-wash buffer, secondary antibody was diluted in Dig-wash buffer (1:100) and incubated with cells for 60min. After washing three times with Dig-wash buffer, cells were re-suspended in 100μL Dig-300 buffer containing 0.58μL hyperactive pG-Tn5, and were incubated at RT for 1h. After washing twice with Dig-300 buffer, 300μL Tagmentation buffer was added and incubated at 37 °C for 1h. The reaction was stopped with 10μL 0.5M EDTA, 3μL 10% SDS and 2.5μL 20 mg/mL Proteinase K for 1h at 50 °C. After DNA extraction with phenol-chloroform and ethanol precipitation, PCR was performed with TruePrep Index Kit V2 for Illumina (Vazyme Biotech, TD202) to amplify the libraries using 16 cycles. All libraries were sequenced by Illumina NovaSeq 6000.