GSM3702000: H3K27ac ChIP Esrrb KO rep1; Mus musculus; ChIP-Seq
Sample information curated by ChIP-Atlas
Antigen
Antigen Class
Histone
Antigen
H3K27ac
Cell type
Cell type Class
Pluripotent stem cell
Cell type
ES cells
NA
NA
Attributes by original data submitter
Sample
source_name
Esrrb -/- embryonic stem cells
chip antibody
H3K27ac
genotype/variation
Esrrb knock-out
Sequenced DNA Library
library_strategy
ChIP-Seq
library_source
GENOMIC
library_selection
ChIP
library_construction_protocol
Chromatin immunoprecipitation (ChIP) was performed as previously described (Frank et al., 2001) with minor modifications outlined below. Chromatin was fixed with 1% Formaldehyde (Sigma-Aldrich) for 10 minutes and sonicated on a bioruptor (Diagenode) to produce fragments of 100-500 bp, and ChIPs performed with Protein G-coupled magnetic Dynabeads (Invitrogen) and the following antibodies : 8 µg MED1 (A300-793A Bethyl Laboratories). The amounts of chromatin (protein) used in each ChIP were as follows: 800 µg (Med1). Following washes of bound DNA-protein complexes, DNA was eluted in 1% SDS and treated with 40 ng/µl RNaseA following 0.2 µg/µl Proteinase K. After phenol/chloroform purification, DNA was then precipitated at -20°C with 20-30 μg GlycoBlue carrier (Invitrogen), 1/10 volumes of 3 M NaAc and 2 volumes of 100% ethanol. Sequencing libraries were prepared using the NEBNext® UltraTM DNA Library Prep Kit and Multiplex Oligos (New England Biolabs) from 5 ng of DNA. Following analysis on an Agilent Bioanalyzer, libraries were pooled and sequenced on an Illumina NextSeq500.