CUT&Tag-seq of PDAC Parental PDO: H3K4me3
CUT&Tag were performedfollowing the protocol of Hyperactive Universal CUT&Tag Assay Kit for Illumina (Vazyme #TD903) to prepare DNA library. Briefly, 5.0104 cells were gently resuspended in NE buffer and incubated on ice for 10 min. The nuclei were isolated and then conjugated to 10 L pre-activated concanavalin A-coated magnetic beads. Then, the nuclei bound to the beads were incubated overnight at 4C in 50 L of Antibody Buffer containing anti-biotin rabbit mAb (1:100 dilution). Subsequently, the nuclei were resuspended in 100 L dig-wash buffer containing antibody (1:100 dilution) to bind with the primary antibody. The nuclei were washed three times with 200 L of dig-wash buffer before being tagmented with the pA/G-Tn5 adapter complex. The tagmented DNA was collected using DNA Extract Beads for library preparation. PCR was performed for library amplification for 15 cycles, and the library was subsequently purified using VAHTS DNA Clean Beads (Vazyme #N411). The tagmented DNA was sequenced using the Illumina NovaSeq 6000 platform.