Antigen

Antigen Class

Histone

Antigen

H3K27ac

Cell type

Cell type Class

Blood

Cell type

Monocytes

MeSH Description

Large, phagocytic mononuclear leukocytes produced in the vertebrate BONE MARROW and released into the BLOOD; contain a large, oval or somewhat indented nucleus surrounded by voluminous cytoplasm and numerous organelles.

Sample

source_name

Monocytes

strain

C57BL/6

tissue

Bone marrow

cell type

Monocytes

chip antibody

anti-H3K27ac

chip antibody vendor

Abcam

Sequenced DNA Library

library_strategy

ChIP-Seq

library_source

GENOMIC

library_selection

ChIP

library_construction_protocol

Chromatin was eluted from the beads, then treated with RNase (Roche) for 30 min at 37°c and with Proteinase K (NEB) for 2 hrs at 37°c. Reverse cross-linking was performed at 65°c overnight. DNA was purified with SPRI beads (Agencourt AMPure XP beads, Beckman Coulter) at 2.3x ratio. Sequential steps of end-repair, A-base addition, adapter-ligation and amplification were performed, with DNA purification with SPRI beads after each step. Following the amplification step, DNA concentration was measured, and equivalent amounts of barcoded ChIPed DNA from each sample were pooled together. An additional size-selection step was performed Libraries were prepared for sequencing using a method developed at Dr. Ido Amit's lab, Weizmann Institute of Science Nat Protoc. 2013 Mar;8(3):539-54. doi: 10.1038/nprot.2013.023. PMID: 23429716

Sequencing Platform

instrument_model

Illumina HiSeq 1500

mm10

Number of total reads

15980646

Reads aligned (%)

96.3

Duplicates removed (%)

9.1

Number of peaks

8498 (qval < 1E-05)

mm9

Number of total reads

15980646

Reads aligned (%)

96.2

Duplicates removed (%)

9.3

Number of peaks

8545 (qval < 1E-05)