GSM4929588: MDA-MB-231 GATA3-flag cells - GATA3 rep1; Homo sapiens; ChIP-Seq
Sample information curated by ChIP-Atlas
Antigen
Antigen Class
TFs and others
Antigen
GATA3
Cell type
Cell type Class
Breast
Cell type
MDA-MB-231
Primary Tissue
Breast
Site of Extraction
Effusion, Pleural
Tissue Diagnosis
Adenocarcinoma
Attributes by original data submitter
Sample
source_name
breast cancer cells
cell line
MDA-MB-231
cell type
triple-negative breast cancer cells
genotype
GATA3-flag wildtype
chip antibody
GATA3 (Abcam, #ab199428-100ul)
Sequenced DNA Library
library_strategy
ChIP-Seq
library_source
GENOMIC
library_selection
ChIP
library_construction_protocol
Lysates were clarified from sonicated nuclei and GATA3-DNA complexes were pulled down with antibody. Libraries were prepared according toTAKARA's instructions accompanying Smarter Thruplex DNAseq Kit. Briefly, in the first step, Template Preparation, the DNA is repaired and yields molecules with blunt ends. In the next step, stem-loop adaptors with blocked 5' ends are ligated with high efficiency to the 5' end of the genomic DNA, leaving a nick at the 3' end. The adaptors can not ligate to each other and do not have single-strand tails, both of which contribute to non-specific background found with many other NGS preparations. In the final step, the 3' ends of the genomic DNA are extended to complete library synthesis, and Illumina-compatible indexes are added through a high-fidelity amplification. Any remaining free adaptors are destroyed. Hands-on time and the risk of contamination are minimized by using a single tube and eliminating intermediate purifications.