|
| Status |
Public on Mar 01, 2010 |
| Title |
Time couse A, 20 minutes |
| Sample type |
genomic |
| |
|
| Channel 1 |
| Source name |
Whole cell extract from 20 minutes after cell cycle release
|
| Organism |
Saccharomyces cerevisiae |
| Characteristics |
strain: W303 background, PSF2-Protein A tagged
|
| Treatment protocol |
alpha-factor was added to a final concentration of 50 nM, and the cells were incubated for another 3 hours prior to release
|
| Growth protocol |
S. cerevisiae strains are from the W303 background and grown in YPD in a 10 L Bioflo 410 fermenter (New Brunswick Scientific) to a density of ~7x106/mL at 30C
|
| Extracted molecule |
genomic DNA |
| Extraction protocol |
Time points were collected in two separate experiments – a) every 15 min after release from the G1 block from 0 min – 105 min and b) every 5 min beginning 20 min after block (through S-phase) from 20 -50 min. For each time-point to be used for ChIP, a 750 mL sample was cross-linked by incubation in 1% formaldehyde at room temperature for 20 min
|
| Label |
Cy3
|
| Label protocol |
Labeling was performed per manufacturers instructions
|
| |
|
| Channel 2 |
| Source name |
IP from 20 minutes after cell cycle release
|
| Organism |
Saccharomyces cerevisiae |
| Characteristics |
strain: W303 background, PSF2-Protein A tagged
|
| Treatment protocol |
alpha-factor was added to a final concentration of 50 nM, and the cells were incubated for another 3 hours prior to release
|
| Growth protocol |
S. cerevisiae strains are from the W303 background and grown in YPD in a 10 L Bioflo 410 fermenter (New Brunswick Scientific) to a density of ~7x106/mL at 30C
|
| Extracted molecule |
genomic DNA |
| Extraction protocol |
Time points were collected in two separate experiments – a) every 15 min after release from the G1 block from 0 min – 105 min and b) every 5 min beginning 20 min after block (through S-phase) from 20 -50 min. For each time-point to be used for ChIP, a 750 mL sample was cross-linked by incubation in 1% formaldehyde at room temperature for 20 min
|
| Label |
Cy5
|
| Label protocol |
Labeling was performed per manufacturers instructions
|
| |
|
| |
| Hybridization protocol |
Agilent Yeast ChIP-on-chip protocol version 9.2
|
| Scan protocol |
Scanned using a PerkinElmer ScanArray Express, with PMT adjusted to 10 points higher than the level where no spots are saturated
|
| Description |
none
|
| Data processing |
Image analysis was performed with Agilent Feature Extraction, converted to GenePix Result (GPR) format and analyzed using Agilent ChIP Analytics
|
| |
|
| Submission date |
Jan 08, 2010 |
| Last update date |
Jan 12, 2010 |
| Contact name |
John Aitchison |
| E-mail(s) |
[email protected]
|
| Phone |
2067321344
|
| Fax |
2067321299
|
| Organization name |
Institute for Systems Biology
|
| Street address |
1441 N34th St
|
| City |
Seattle |
| State/province |
WA |
| ZIP/Postal code |
98103 |
| Country |
USA |
| |
|
| Platform ID |
GPL4131 |
| Series (1) |
| GSE19818 |
Saccharomyces cerevisiae GINS complex progression (Psf2) determined ChIP-chip time course through the cell cycle |
|
