vmrcrcw_igg
Homo sapiens
Protocols: 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.
ENA-FIRST-PUBLIC
2019-06-28T04:03:10Z
ENA-LAST-UPDATE
2019-03-22T12:23:56Z
External Id
SAMEA5529085
INSDC center name
Novartis Institutes for Biomedical Research
INSDC first public
2019-06-28T04:03:10Z
INSDC last update
2019-03-22T12:23:56Z
INSDC status
public
Submitter Id
E-MTAB-7812:vmrcrcw_igg
broker name
ArrayExpress
cell line
VMRC-RCW
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:vmrcrcw_igg
scientific_name
Homo sapiens