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| Status |
Public on Apr 25, 2019 |
| Title |
Genome-wide binding of the meiosis specific Rec8 cohesin subunit in metaphase I |
| Organism |
Saccharomyces cerevisiae |
| Experiment type |
Genome binding/occupancy profiling by high throughput sequencing
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| Summary |
ChIP-seq profile of Rec8 binding sites arrested in metaphase I by depletion of the APC/C activator CDC20. The aim of this experiment is to identify Rec8 binding sites in metaphase I.
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| Overall design |
Diploid S. cerevisiae SK1 cells were constructed containing Rec8-3Ha in an pGAL1-NDT80 pGPD1-GAL4.ER pCLB2-CDC20 background. Cells were allowed to sporulate in sporulation media for 5 hours before addition of β-estradiol to release from prophase arrest. Cells were grown for 75min to arrest in metaphase I after which formaldehyde was added to the cultures and cells were fixed for 30min. We chose this double-arrest strategy (with a prophase I arrest followed by a metaphase I arrest) to ensure efficient arrest in metaphase I, which is less robust in cells without Spo13. Samples were subsequently processed according to the standard lab ChIP-Seq protocol.
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| Contributor(s) |
Galander S, Marston A |
| Citation(s) |
31031198 |
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| Submission date |
Dec 10, 2018 |
| Last update date |
Jun 21, 2019 |
| Contact name |
Daniel Robertson |
| E-mail(s) |
daniel.robertson@ed.ac.uk
|
| Organization name |
University of Edinburgh
|
| Department |
Discovery Research Platform for Hidden Cell Biology
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| Lab |
Bioinformatics Core
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| Street address |
2.28 Michael Swann Building, Kings Buildings, Mayfield Road
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| City |
Edinburgh |
| State/province |
Midlothian |
| ZIP/Postal code |
EH9 3JR |
| Country |
United Kingdom |
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| Platforms (1) |
| GPL22715 |
Illumina MiniSeq (Saccharomyces cerevisiae) |
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| Samples (2) |
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| This SubSeries is part of SuperSeries: |
| GSE112217 |
Reductional meiosis I chromosome segregation is established by coordination of key meiotic kinases |
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| Relations |
| BioProject |
PRJNA509161 |
| SRA |
SRP173222 |
