Genome-wide Cohesin Binding Profiling

At the indicated times, aliquots of the cultures were harvested and processed for ChIP of the cohesin complex as described [24 (link)]. Antibodies used for ChIP were α-Pk (clone SV5-Pk1, AbD Serotec), α-myc (clone 9E10) and α-HA (clone 12CA5). Processed chromatin immunoprecipitates and input DNA samples were hybridized to Affymetrix GeneChip S. cerevisiae Tiling 1.0R arrays. Presented is the genome-normalized ratio of the hybridization signals of the chromatin immunoprecipitate over the input DNA. Each bar in the bar graphs represents the average of 25 oligonucleotide probes within neighbouring 125 bp windows. For the line graphs, a smoothed moving average of 200–500 bp is shown. The microarray data are available from the GEO database under the accession number GSE80464. The quantitative analysis of cohesin binding to individual loci was performed as described, using previously described qPCR primer pairs [30 (link)], as well as primer pairs flanking the GAL2 locus, listed in the electronic supplementary material, table S2.