title: Illumina HiSeq 4000 sequencing; ChIP-seq of H3K27ac and H3K27me3 in HSJD-DIPG-007 cell line
design: ChIP-seq of H3K27ac and H3K27me3 in HSJD-DIPG-007 cell line
library_name: HSJD-DIPG-007_H3K27me3_2_s
library_construction_protocol: Cells were crosslinked in 1% formaldehyde followed by quenching with Glycine (Final concentration 125 mM). Samples were then lysed and sonicated for 20 cycles (30 seconds on/ 30 seconds off) (Bioruptor pico, Diagenode). Precleared chromatin was incubated with 1µg antibody for H3K27ac (196-050, Diagenode), H3K27me3 (C15410195, Diagenode), and rabbit IgG (C15410206, Diagenode). Protein A sepharose beads were used to pull down the antibody-chromatin complex and samples were then de-crosslinked and DNA was extracted.  DNA was extracted using phenol/chlorofom/Isoamylic alcohol (25:24:1). Aqueous phase was collected and DNA was precipitated using 100% ethanol followed by washing with 70% ethanol and resuspension in water.  Microplex Library preparation kit v2 (diagenode) was used to prepare libraries from ChIP DNA according to manufacturer's instructions.
attributes.tag: Experimental Factor: immunoprecipitate
attributes.value: H3K27me3