GSM3752934: 7-19T-Young-Input; Homo sapiens; ChIP-Seq
Sample information curated by ChIP-Atlas
Antigen
Antigen Class
Input control
Antigen
Input control
Cell type
Cell type Class
Neural
Cell type
Lateral temporal lobe
NA
NA
Attributes by original data submitter
Sample
source_name
Frozen postmortem brain tissue
biomaterial_provider
CNDR brain bank (Penn)
disease state
Young
tissue
Lateral temporal lobe
chip antibody
input
Sequenced DNA Library
library_strategy
ChIP-Seq
library_source
GENOMIC
library_selection
ChIP
library_construction_protocol
Nuclei were isolated from 200 mg frozen brain tissue by dounce homogenization in nuclei isolation buffer followed by ultracentrifugation on a 1.8 M sucrose cushion. Nuclei pellets were resuspended and crosslinked in 1% formaldehyde for 10 min at RT, quenched with 125 mM glycine for 5 min and washed twice in cold PBS. 2 x 106 nuclei were lysed in nuclei lysis buffer and chromatin was sheared using a Covaris S220 sonicator to a final ~250 bp. Equal aliquots of sonicated chromatin were used per immunoprecipitation (IP) reaction with antibody (either H3K27ac, or H3K9ac or H3K122ac or H3K4em1) preconjugated to Protein G Dynabeads. 10% of each IP was used for the Inputs. ChIP reactions were incubated overnight at 4°C followed by ChIP washes and DNA was eluted in elution buffer (1% SDS, 50 mM Tris-HCl pH 8, 10 mM EDTA) at 65°C. Eluted DNA from ChIP and Input samples was reverse crosslinked and purified using PCR columns (Qiagen). Sequencing libraries for ChIP experiments were prepared using NEBNext Ultra II reagents (New England Biolabs). All ChIP samples and Input were single-end sequenced on an Illumina NextSeq 500.