GSM4282544: 293 KO Chip1 Hyp ; Homo sapiens; ChIP-Seq
Sample information curated by ChIP-Atlas
Antigen
Antigen Class
TFs and others
Antigen
HIF1A
Cell type
Cell type Class
Kidney
Cell type
293
Primary Tissue
Kidney
Tissue Diagnosis
Normal
Attributes by original data submitter
Sample
source_name
HEK293T K.O. PARP-1 cells
cell line
HEK293T
cell type
human embryonic kidney cells
passages
5 to 10
genotype/variation
PARP-1 K.O.
condition
Hypoxia
chip antibody
Anti-HIF-1 alpha antibody - ChIP Grade (ab2185)
Sequenced DNA Library
library_strategy
ChIP-Seq
library_source
GENOMIC
library_selection
ChIP
library_construction_protocol
Cellular lysates were treated with micrococcal nuclease in order to digest genomic DNA. HIF-1α-DNA complexes were isolated with antibody. Library preparation and Illumina sequencing were performed by the IPBLN Genomics Core Facility (CSIC, Granada, Spain). ChIP-seq libraries were generated from 30 ng of DNA using the NEBNext Ultra II DNA library preparation kit for Illumina (NEB), and following manufacturer´s instructions with some modifications. A 10-fold dilution of adaptor was used in ligation reaction and no size selection was applied to the adaptor-ligated DNA clean up step, as recommended for Micrococcal Nuclease digested chromatin ChIP assays. A total of 5 cycles of amplification were used to complete adaptor sequences and introduce unique indices for sample multiplexing. Final libraries were quantified on the Qubit® fluorometer (Thermo) and run on a Bioanalyzer HS DNA chip to verify quality and size distribution. Equimolecular library pool was prepared, then diluted and denatured as recommended by Illumina NextSeq 500 library preparation guide. The 75 nt single-end sequencing was conducted on a NextSeq 500 sequencer (Illumina). An average of 24 million reads per sample was obtained.