Illumina HiSeq 2500 paired end sequencing; ChIP-seq for PAX8 and histone modifications in Renal Cell Carcinoma cell lines.
Sample information curated by ChIP-Atlas
Antigen
Antigen Class
No description
Antigen
NA
Cell type
Cell type Class
Kidney
Cell type
769-P
Primary Tissue
Kidney
Tissue Diagnosis
Adenocarcinoma
Attributes by original data submitter
Sample
ENA first public
2019-06-28
ENA last update
2019-03-22
ENA-CHECKLIST
ERC000011
External Id
SAMEA5529068
INSDC center alias
Novartis Institutes for Biomedical Research
INSDC center name
Novartis Institutes for Biomedical Research
INSDC first public
2019-06-28T04:03:10Z
INSDC last update
2019-03-22T12:23:55Z
INSDC status
public
Submitter Id
E-MTAB-7812:769p_h3k4me1
broker name
ArrayExpress
cell line
769P
cell type
kidney cell
common name
human
developmental stage
adult
disease
clear cell renal carcinoma
genotype
PAX8 doxycycline inducible KD
organism part
kidney
sample name
E-MTAB-7812:769p_h3k4me1
Sequenced DNA Library
library_name
769p_h3k4me1_p
library_strategy
ChIP-Seq
library_source
GENOMIC
library_selection
ChIP
library_construction_protocol
Chromatin immunoprecipitation was performed as previously described (Galli et al., 2015). Briefly, 8*15 cm dishes of cells were cross-linked with 1% formaldehyde at room temperature for 10 minutes. The cross-linking was quenched by fresh glycine at a final concentration of 0.125 M. The cells were then harvested and lysed in SDS lysis buffer to collect the nuclei. Nuclei suspension was sonicated using a Diagenode Bioruptor (14 cycles 30\" ON and 30\" OFF using setting High) to obtain fragments of 250bp average size. Appropriate amount of chromatin (100ug for PAX8 or 10ug for histone modification) was incubated with the indicated antibodies overnight and complexes retrieved using Protein G coupled Dynabeads. After extensive washes, DNA was reverse crosslinked and purified using AmpPure XP beads. ChIP-seq libraries were constructed using Nugen Ovation® Ultralow System V2 according to manufacturer's instructions.