Sample information curated by ChIP-Atlas

Antigen

Antigen Class
Histone
Antigen
H3K27ac

Cell type

Cell type Class
Neural
Cell type
Cerebellum neurosphere
NA
NA

Attributes by original data submitter

Sample

source_name
Neurosphere-from cerebellum
cell type
Neurosphere
tissue
neurosphere derived from cerebellum
genotype
wildtype
chip antibody
H3K27ac (Abcam, cat# ab4729)

Sequenced DNA Library

library_strategy
ChIP-Seq
library_source
GENOMIC
library_selection
ChIP
library_construction_protocol
ChIP assays were performed as a modification of the previously described methods (Dhar et al., 2016; Dhar et al., 2012). In brief, cerebellar (4-month-old) tissues were isolated. DNA was purified from chromatin immunoprecipitates for MLL4 or IgG using the phenol/chloroform extraction. IgG was used as a negative control for ChIP. Then, DNA was amplified by quantitative PCR and normalized to input. To compare H3K4me1, H3K27ac, and H3K4me3 levels between Mll4f/f and Mll4 BSKO cerebellum, ChIP with reference exogenous genome (ChIP-Rx) was carried out as previously described (Li et al., 2016; Orlando et al., 2014). In brief, two biological replicates for Mll4f/f and Mll4 BSKO cerebellum were used for ChIP-Rx experiments. Drosophila melanogaster chromatin and Drosophila-specific H2Av antibody were added to each ChIP assay as a minor fraction. The following components for ChIP-Rx were purchased from Active Motif: the spike-in chromatin (53083), Drosophila-specific H2Av antibody (61686), spike-in ChIP positive control primer set (71037), and negative control primer set (71028). The immunoprecipitated DNAs were sequenced by Next-Generation Sequencing (NGS) (UCI GHTF). ChIP DNA libraries were prepared for sequencing using standard Illumina protocols.

Sequencing Platform

instrument_model
Illumina HiSeq 2000

mm10

Number of total reads
7174342
Reads aligned (%)
87.1
Duplicates removed (%)
39.9
Number of peaks
6179 (qval < 1E-05)

mm9

Number of total reads
7174342
Reads aligned (%)
86.5
Duplicates removed (%)
40.3
Number of peaks
6199 (qval < 1E-05)

Base call quality data from DBCLS SRA