ChIP-Atlas: SRX2409209

Antigen

Cell type Class: Others
Cell type: Dura Mater
MeSH Description: The outermost of the three MENINGES, a fibrous membrane of connective tissue that covers the brain and the spinal cord.

library_strategy: ChIP-Seq
library_source: GENOMIC
library_selection: ChIP
library_construction_protocol: 10 to 15 frozen sections of each tumor block or dura sample were collected for each experiment. Tissue was crosslinked with 1% formaldehyde, quenched with glycine, and washed with PBS. Nuclei were extracted by dounce homogenization and resuspended in nuclear lysis buffer containing 0.3% SDS. Chromatin was sheared by sonication with a Q800R2 Sonicator by QSonica (60 minutes total, amplitude 30, 10 second pulses, 10 second rest). Soluble chromatin was incubated with magnetic beads coated with either H3K27ac antibody (ab4729) overnight at 4°C. Chromatin was precipitated using a magnet, washed extensively, and eluted with TE+1% SDS. Crosslinks were reversed, and samples were purified. Prior to sequencing, the enrichment of each sample was calculated using qPCR and well-established control primers for H3K27ac.. Samples were subjected to standard Illumina paired-end multiplexed library construction. H3K27ac ChIP and input samples were sequenced for each tumor (1x75bp, HiSeq 2000).