GSM8354197: dCas9-DNMT3A_ChIP_CTRL_r1; Homo sapiens; ChIP-Seq
ChIP experiments were performed as previously described (Sardina et al. Cell Stem Cell. 2018). Briefly, 30 million cells were cross-linked with 1% formaldehyde (Sigma).  Chromatin was sheared to 100-300bp fragments and conjugated with 10ug of anti-Cas9 antibody (Active Motif Cat #61757) rotating ON at 4ºC.Finally, DNA was purified by phenol: chloroform:isoamyl alcohol (25:24:1) extraction and ethanol precipitated. For ChIP-seq, IP and Input DNA samples were quantified with Agilent 2100 before library preparation. BGI Genomics (Hong Kong, China) performed the library construction and sequencing process. For ligation, DNA-end repair and 3'-dA overhang was used. Then PCR amplification and size selection was performed. Library quality control and circularization was performed prior amplification to make DNA nanoball (DNB). Sequencing was done on the DNBSEQ platform using 50-nucleotide paired-end mode.