E-MTAB-2701:IgG_control_R1
Homo sapiens
Protocols: After reverse cross-link and protein digestion with Proteinase K (Roche), DNA was purified using Qiaquick, PCR purification kit (Qiagen) following the manufacturer's instructions. The stable cell lines were grown in DMEM supplement with 10% foetal bovine serum. Media contained 100 micro-g/ml of Hygromycin. Expression of ZMYM2(WT) or ZMYM2(SIM2mut) was induced by treating the cells for 48 hours with 100 ng/ml [for ZMYM2(WT)] or 50 ng/ml [for ZMYM2(SIM2mut)] doxycycline (Sigma). Expression of FOXO3 was induced by treating the cells for 48 hours with 1 ng/ml doxycycline (Sigma). Cells were double cross-linked with dimethyl adipimidate (DMA) for 20 minutes and subsequently with Formaldehyde for 10 minutes. ChIP-seq was performed as described in Schmidt et al., 2009, using 60 x 106 U2OS-ZMYM2(WT) or U2OS-ZMYM2(SIM2mut). Immuno-precipitation was performed overnight at 4 degrees C by incubating the shared DNA-protein complex with 6 ug of anti-Flag (Sigma) or control IgG (Millipore) antibodies previously coupled to Dynabeads protein G (Invitrogen). Schmidt D, Wilson MD, Spyrou C, Brown GD, Hadfield J, Odom DT. (2009) ChIP-seq: using high-throughput sequencing to discover protein-DNA interactions. Methods 48: 240-248. Note: paired-end sequencing libraires were created in this experiment but only the forward (R1) reads are available in this submission. The 'LIBRARY_LAYOUT' attribute is therefore set to be 'SINGLE' rather than 'PAIRED'.
ENA checklist
ERC000011
INSDC center alias
University of Manchester
INSDC center name
University of Manchester
INSDC first public
2016-06-27T11:01:18Z
INSDC last update
2018-03-08T18:52:56Z
INSDC status
public
SRA accession
ERS496031
Sample Name
ERS496031
alias
E-MTAB-2701:IgG_control_R1
broker name
ArrayExpress
cell line
U2OS
cell type
osteosarcoma
organism
Homo sapiens
sex
female
title
IgG_control_R1