|
| Status |
Public on Sep 30, 2007 |
| Title |
GAL-Scc1-HA3 distribution (noc-loaded; +NOC) |
| Sample type |
genomic |
| |
|
| Source name |
Chromatin-immunoprecipitated DNA
|
| Organism |
Saccharomyces cerevisiae |
| Characteristics |
Strain 3267 = Scc1-9PK-TRP, leu:GAL-SCC1-HA3:LEU2, MET-HA3-CDC20-URA3
|
| Treatment protocol |
Cells were synchronized in G1 with α-factor and released into HU arrest in early S-phase. From there cells were released into metaphase arrest by Cdc20 depletion in the presence of nocodazole. Loading of ectopic, HA epitope-tagged cohesin was induced by induction of the GAL1 promoter for 1 hour during the time in the metaphase arrest (noc-loaded).
|
| Extracted molecule |
genomic DNA |
| Extraction protocol |
Extraction was performed as previously described (Katou et al. 2003).
Reference:
Katou Y, Kanoh Y, Bando M, Noguchi H, Tanaka H, Ashikari T, Sugimoto K and Shirahige K. (2003) S-phase checkpoint proteins Tof1 and Mrc1 form a stable replication-pausing complex. Nature 424: 1078-1083.
|
| Label |
biotin-N6-ddATP
|
| Label protocol |
DNA was labeled with biotin-N6-ddATP using terminal transferase TdT (Roche, 400U/µl).
|
| |
|
| Hybridization protocol |
Hybridization was performed as previously described (Katou et al. 2003). Samples were hybridized for 16 hours at 42ºC in a hybridization oven (GeneChip Hybridization Oven 640, Affymetrix).
Reference:
Katou Y, Kanoh Y, Bando M, Noguchi H, Tanaka H, Ashikari T, Sugimoto K and Shirahige K. (2003) S-phase checkpoint proteins Tof1 and Mrc1 form a stable replication-pausing complex. Nature 424: 1078-1083.
|
| Scan protocol |
The washing and scanning protocol, provided by Affymetrix, was performed automatically on a fluidics station (GeneChip fluidics station 450, Affymetrix).
|
| Description |
GAL-Scc1-HA3 distribution (noc-loaded) on chromosome VI in metaphase-arrested cells in the presence of nocodazole. [Figure 4, 3rd panel]
|
| Data processing |
Data were processed following the protocol provided by Affymetrix (GCOS ver 1.4).
|
| |
|
| Submission date |
Aug 01, 2007 |
| Last update date |
Aug 14, 2011 |
| Contact name |
Maria Ocampo-Hafalla |
| Organization name |
Cancer Research UK London Research Institute
|
| Lab |
Chromosome Segregation
|
| Street address |
44 Lincoln's Inn Fields
|
| City |
London |
| ZIP/Postal code |
WC2A 3PX |
| Country |
United Kingdom |
| |
|
| Platform ID |
GPL347 |
| Series (1) |
| GSE8661 |
Displacement and re-accumulation of centromeric cohesin during transient pre-anaphase centromere splitting |
|
