GSM7816257: translocation_HSPC_H3K4me1_3; Homo sapiens; OTHER
Cell permeabilization of native cells and tagmentation reaction with pA-Tn5 transposome-antibody complex for "Antibody-guided chromatin tagmentation (ACT-seq)" according to Carter et al (PMID: 31431618). For normalization of sequence reads between biological replicates, ~4,000 permeabilized nuclei of yeast Saccharomyces cerevisiae, prepared according to (PMID: 25827087) and incubated with pA-Tn5ome-ab complex targeting yeast H2B, were spiked into each mix of cells and pA-Tn5ome-ab complex.  Tagmented DNA was purified with MinElute (Qiagen) columns (ACTseq) according to the provider's instruction. Libraries were prepared according to Weichenhan et al. (PMID: 29224141). Briefly, tagmented DNA was amplified by PCR under real-time conditions using a universal Illumina forward and a barcoded reverse primer and the NEBNext High Fidelity 2x PCR Master-Mix (NEB), and amplfication was monitored by inspecting real time the amplification curves. DNA was HighPrep bead-purified, Qubit (Invitrogen) quantitated and sized by TapeStation (Agilent) profiling.