GSM2150985: SVZ H4K20me3 ChIP-Seq; Papio anubis; ChIP-Seq
We manually conjugated antibody against cell type markers, such as GFAP, Nestin, Vimentin, PSA-NCAM, or Doublecortin to Dynabeads. We then used the Dynabeads-conjugated antibody to purify cells dissociated from SVZ microdissection. Briefly, cells from fresh dissected baboon SVZ were immediately dissociated with Accutase, equilibrated in binding buffer containing phosphate-buffered saline (PBS), 0.05% TritonX-100 (or saponin, detergent choice depends upon antibody), and subsequently subjected to Dynabeads-conjugated antibody purification. After elution with high salt and pH-gradient buffer, the purified populations were crosslinked in 1.1% formaldehyde before chromatin shearing by Diagenode Bioruptor. The resulting sheared chromatin fragments in a size range between 200 to 500 base pairs were incubated with Protein A Dynabeads (Life Technology Dynabeads protein A) conjugated to H3K27me3 antibody (Active Motif #39155; 1:1000) or H4K20me3 antibody (Upstate #07-463; 1:1000) overnight. For normalization, the aliquot of sheared chromatin fragments were incubated with antibody against total histone 3-conjugated Protein A Dynabeads (unmodified H3 antibody, Millipore #05-499; 1:1000) or total H4 (Cell Signaling #2592; 1:1000). Enriched chromatin was eluted and de-crosslinked, prior to library preparation with TruSeq ChIP sample prep kit (Illumina #IP-202-2012 and #IP-202-1024) and deep sequencing to > 200 million tags on an Illumina HiSeq2000 sequencer. Libraries were prepared according to instructions for Illumina Library Kit, Illumina  TruSeq v2
GEO Accession
GSM2150985