Illumina HiSeq 2500 paired end sequencing; SUMO2 ChIP-seq in U2OS cells
SUMO2 ChIP-seq in U2OS cells
The U2OS cells were grown in DMEM supplement with 10% foetal bovine serum.  Cells were  cross-linked with  Formaldehyde for 10 minutes. 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. ChIP-seq was performed as described in Schmidt et al., 2009, using 3  x 10^6 U2OS. Immuno-precipitation was performed overnight at 4° C by incubating the shared DNA-protein complex with3 micro-gram of anti-SUMO2 (Life technologies) antibody previously coupled to Dynabeads protein A (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. Libraries were generated following manufacturer's protocol.
UNIVERSITY OF MANCHESTER
E-MTAB-3084:Input DNA
University of Manchester
E-MTAB-3084:Input DNA