GSM6008197: HEK293T, Cut&Tag, siDot1L, OGT, rep2; Homo sapiens; OTHER
GSM6008197
HEK293T cells were harvested and washed three times by PBS Cut&Tag experiments were performed with Hieff In-Situ DNA Binding Profiling Library Prep Kit (Yeasen, 12598ES48) following manufacturer's protocols. Briefly, for each sample, 5x104 cells were harvested and washed three times by PBS, resuspend the cells with cell wash buffer gently, and incubate with the concanavalin A coated magnetic beads previously washed by ConA bind buffer. Then concanavalin A beads coupled cells were collected and incubated with primary antibody Rabbit IgG (Abcam) or Dot1L(CST) or OGT(Proteintech) at 4 °C overnight. Incubate the secondary antibody with the concanavalin A beads coupled cells at room temperature 1 h. Collecting the ConA beads coupled cells and the pA/G-Transposome Mix was added. And standard tagmentation and amplification were performed. Amplified DNA libraries were purified and shipped for next-generation sequencing (NGS) sequencing.