GSM4349539: PDX34 CC SJBALL205 GATA3 ChIP-seq; Homo sapiens; ChIP-Seq
Sample information curated by ChIP-Atlas
Antigen
Antigen Class
TFs and others
Antigen
GATA3
Cell type
Cell type Class
Unclassified
Cell type
Unclassified
NA
NA
Attributes by original data submitter
Sample
source_name
Tissue
tissue
patient-derived xenografts
chip-seq antibody
GATA3 antibody(Santa Cruz, sc268)
Sequenced DNA Library
library_strategy
ChIP-Seq
library_source
GENOMIC
library_selection
ChIP
library_construction_protocol
10 M cells are fixed with 1% FA for 15 mins in room temperature, then stoped with Glycine. Chromatin was extracted with cell lysis buffer (1% SDS, 50 mM Tris-HCl pH 8, 20 mM EDTA). 100 ug chromatin was sonicated to 100-300 bp by Covaris E220, and 5 ug chromatin was used as input. Bead-antibody complex was prepared by incubating 11 ul of sheep anti-mouse IgG dynabeads (ThermoFisher, 11201D) with anti-GATA3 (Santa Cruz, sc268) at 4°C for 4 hours with shaking. Then fragmented chromatin was incubated with bead-antibody complex overnight with shaking followed by stringent wash and elution. To prepare the library, eluted chromatin was reverse crosslinked and purified by phenol-chloroform extraction. Then DNA was end-repaired by END-IT DNA end-repair kit (Epicentre, ER81050) according to kit's protocol, added A using Klenow fragment (3'->5' exo-) (NEB, M0212S), ligated with Illumina TruSeq adaptor (Illumina, FC-121-3001) and subsequently amplified by PCR (Roche, kk2601). The quality and quantity of all the libraries were checked using BioAnalyzer High Sensitivity DNA Kit (Agilent). The libraries were sequenced on Illumina HiSeq platform.