Alvocidib ChIP
Homo sapiens
Protocols: CrizR1 human cells are derived from the parental NCI-H3122 (CVCL_5160) and are resistant to crizotinib. CrizR1 cells were grown in 10cm2 dishes to 80% confluence and treated with DMSO, 200nM alvocidib or 100nM THZ1 for 6 hrs. Afterwards, cells were cross-linked in 1% formaldehyde for 10 mins at room temperature. After washing with ice-cold PBS, cells were collected by scraping in ice-cold PBS supplemented with protease inhibitors and genomic DNA isolated as follows. The protocol by (Nelson et al., 2006; https://doi.org/10.1038/nprot.2006.27) was followed to isolate chromatin. Briefly, cell pellets were sonicated for 25 cycles, with a 30s on, 45s off protocol. Then the input was reversed crosslinked and analysed on a gel to confirm efficient sonication. Input was then incubated O/N with Protein G beads bound to 10ug of total RNA polymerase II antibody (Diagenode, #C15200004) per sample. The chromatin was washed and eluted as per the cited protocol. The quality of the DNA was assessed using 2100 Agilent Bioanalyzer. Then, purified chromatin was used as input to generate PCR-amplified libraries using the Diagenode Microplex kit according to the manufacturer's instructions.
ENA-FIRST-PUBLIC
2020-05-13T17:05:17Z
ENA-LAST-UPDATE
2019-09-25T16:27:01Z
External Id
SAMEA5975616
INSDC center name
UNIVERSITY OF MANCHESTER
INSDC first public
2020-05-13T17:05:17Z
INSDC last update
2019-09-25T16:27:01Z
INSDC status
public
Submitter Id
E-MTAB-8380:Alvocidib ChIP
broker name
ArrayExpress
cell line
CrizR1
common name
human
developmental stage
adult
disease
lung adenocarcinoma
organism part
lung
parental cell line
NCI-H3122
phenotype
resistant to crizotinib
sample name
E-MTAB-8380:Alvocidib ChIP
scientific_name
Homo sapiens