GSM1227212: U-2932 ChIPSeq; Homo sapiens; ChIP-Seq
Sample information curated by ChIP-Atlas
Antigen
Antigen Class
TFs and others
Antigen
STAT3
Cell type
Cell type Class
Blood
Cell type
U-2932
Primary Tissue
Blood
Site of Extraction
Ascites
Tissue Diagnosis
Lymphoma B-cell
Attributes by original data submitter
Sample
source_name
B cells
disease state
diffuse large B cell lymphoma (DLBCL)
dlbcl subtype
ABC
cell line
U-2932
chip antibody
STAT3 (Santa Cruz, sc-482X)
Sequenced DNA Library
library_strategy
ChIP-Seq
library_source
GENOMIC
library_selection
ChIP
library_construction_protocol
Crosslinked cell pellets were dounce homogenized to collect nuclei. Nuclear lysate was sonicated, then immunoprecipitated. Antibody-protein-DNA complexes were collected using Protein A agarose beads. Libraries were prepared according to Illumina's instructions using non-Illumina enzymes and kits. Briefly, DNA was end-repaired using a combination of T4 DNA polymerase, E. coli DNA Pol I large fragment (Klenow polymerase) and T4 polynucleotide kinase (End-It Repair Kit, Epicenter-Illumina). The blunt ends were treated with Klenow fragment (32 to 52 exo minus) and dATP to yield a protruding 3- 'A' base for ligation of Illumina's adapters. DNA was then gel purified and size selected for 150-300 bp fragments to exclude unligated adapters, PCR amplified with Illumina primers using Phusion high-fidelity DNA polymerase for 15 cycles, and size selected again from an agarose gel for 150-300 bp fragments. The purified DNA was captured on an Illumina flow cell for cluster generation. Libraries were sequenced on the Genome Analyzer IIx following the manufacturer's protocols.