GSM5342482: I-BET151-BRD4_rep1; Homo sapiens; OTHER
CUT&Tag assay was performed as follows. First, ConA Beads (CST, #93569) were pipetted into a new tube, 10 μl per sample, and washed by Activation Buffer. 1 × 10^5 cells on d7 were harvested and washed by Wash Buffer. The cells were mixed with ConA Beads and incubated with Anti-Brd4 antibody (Abcam) (1:100) overnight at 4°C. Donkey anti-rabbit lgG antibody (Invitrogen) was diluted at 1:100 and added into the samples. then, the samples were incubated for 1 h at RT. After washing away the unbounded secondary antibody, Hyperactive pG-Tn5 Transposase (Vazyme) was added and incubated with cells for 1 h at RT. The samples were washed by Dig-med Buffer and followed with tagmentation by Tagmentation Buffer. Next, DNA was isolated by phenol-chloroform and dissolved in TE-buffer. The standard processing was performed using TruePrep DNA Library Prep Kit V2 for Illumina (Vazyme) and TruePrep Index Kit V2 for Illumina ® (Vazyme TD202). The library concentration was measured using Qubit dsDNA BR Assay Kit (Invitrogen) by Qubit (Invitrogen).
GEO
GSM5342482