GSM2966296: AR shKDM3A ChIPSeq; Homo sapiens; ChIP-Seq
Sample information curated by ChIP-Atlas
Antigen
Antigen Class
No description
Antigen
NA
Cell type
Cell type Class
No description
Cell type
NA
NA
NA
Attributes by original data submitter
Sample
Sequenced DNA Library
library_strategy
ChIP-Seq
library_source
GENOMIC
library_selection
ChIP
library_construction_protocol
Cells were crosslinked using 1% formaldehyde for 10 min at 298 K. Formaldehyde was removed and cells were incubated with 125 mM glycine for 5 min at 298 K. Nuclear extracts were collected and sonicated to obtain 300 bp chromatin fragments using the Covaris S2 ultrasonicator (Covaris, Woburn, MA). 100 µg of chromatin was incubated with 5 µg of AR (PG21, 06-680, Sigma EMD Millipore, Darmstadt, Germany), 5 µg of KDM3A (A301-538A, Bethyl Laboratories, Montgomery, TX), 2 µg of H3K9me1 (ab9045, Abcam, Cambridge, MA), or 2 µg of H3K9me2 (07-441, Sigma EMD Millipore, Darmstadt, Germany) antibodies overnight at 277 K followed by incubation with 30 µl of protein A/G beads for 4 hours. After four washes, crosslinking was reversed. Chromatin was digested with ribonuclease A followed by proteinase K. Then the DNA was purified using spin columns. The size of the DNA was confirmed by a bioanalyzer (Agilent Biotechnologies, Savage, MD). The purified DNA library was sequenced using an Illumina HighSeq2000 at the Sanford-Burnham Medical Research Institute at Lake Nona, National Genome Library Core Facility.