ChIP-Atlas: SRX1177279

Antigen

Cell type Class: Neural
Cell type: Prefrontal Cortex
MeSH Description: The rostral part of the frontal lobe, bounded by the inferior precentral fissure in humans, which receives projection fibers from the MEDIODORSAL NUCLEUS OF THE THALAMUS. The prefrontal cortex receives afferent fibers from numerous structures of the DIENCEPHALON; MESENCEPHALON; and LIMBIC SYSTEM as well as cortical afferents of visual, auditory, and somatic origin.

library_strategy: ChIP-Seq
library_source: GENOMIC
library_selection: ChIP
library_construction_protocol: Chromatin immunoprecipitation (ChIP) was performed as previously described (Heintzman et al., 2007). Briefly, 10-20 million of cultured cells and ~0.2 g of frozen tissue samples were cross-linked in 1% formaldehyde at room temperature for 10 and 25 minutes, respectively. Cross-linked samples were sonicated until chromatin fragments were of 200-600 bp in length. Chromatin immunoprecipitation was carried out using 3-5 µg of antibody and 200 µg of chromatin preparation in RIPA buffer at 4 degree overnight. After five washes in RIPA and one wash with 1xTE buffer, chromatins were eluted and reverse cross-linked at 65 oC overnight. DNA was purified after phenol extraction. Libraries were prepared according to Illumina's instructions, using TruSeq DNA Sample Preparation Kits v2, Set A (Cat# FC-121-2001). 4-12 library preps were mixed for multiplexed single-read sequencing using Illumina Hi-seq 2000 (Illumina).