|
| Status |
Public on Jun 11, 2011 |
| Title |
Glucose cln1,2,3∆ MET3pr-CLN2 timepoint min 5 |
| Sample type |
RNA |
| |
|
| Channel 1 |
| Source name |
lack of G1 cyclin synchronized cells 0 minute
|
| Organism |
Saccharomyces cerevisiae |
| Characteristics |
strain: W303
genotype: cln1,2,3∆ MET3pr-CLN2
time: 0 min
media: glucose
|
| Treatment protocol |
Then, cells were transferred to SCD-methionine (lacking methionine) and let them wait for 120 minutes to arrest. Cells are released by adding .25Xmethionine which leads to express CLN2 at physiological level under MET3 promoter. Then samples are drawn every 5 minutes for 60 minutes. RNAs are hybridized on the first time point, i.e. 0 minute
|
| Growth protocol |
Yeast cells were grown over night in SCD complete media, in 300C.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
RNAs were isolated by using Ambion Yeast RiboPure Kit,
|
| Label |
Cy5
|
| Label protocol |
RNAs were labeled by Aglient “Quick Amp Labeling Kit Two-color”
|
| |
|
| Channel 2 |
| Source name |
lack of G1 cyclin synchronized cells
|
| Organism |
Saccharomyces cerevisiae |
| Characteristics |
media: glucose
genotype: cln1,2,3∆ MET3pr-CLN2
time: 5 min
strain: W303
|
| Treatment protocol |
Then, cells were transferred to SCD-methionine (lacking methionine) and let them wait for 120 minutes to arrest. Cells are released by adding .25Xmethionine which leads to express CLN2 at physiological level under MET3 promoter. Then samples are drawn every 5 minutes for 60 minutes. RNAs are hybridized on the first time point, i.e. 0 minute
|
| Growth protocol |
Yeast cells were grown over night in SCD complete media, in 300C.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
RNAs were isolated by using Ambion Yeast RiboPure Kit,
|
| Label |
Cy3
|
| Label protocol |
RNAs were labeled by Aglient “Quick Amp Labeling Kit Two-color”
|
| |
|
| |
| Hybridization protocol |
1 Equilibrate water bath to 60°C.2.For 8x15K microarray format, add cyanine 3-labeled, linearly amplified cRNA 300 ng cyanine 3-labeled, linearly amplified cRNA 300ng, 10X GEx Blocking Solution 5ul, Nulease free H2O bring Vol to 24ul, 25X Fragmentation Buffer 1ul, Total Vol 25ul, 3. Incubate at 60°C for exactly 30 minutes to fragment RNA. 4. Add 25ul 2x GEx Hybridization Buffer HI-RPM to the 8X15K. 5. Place sample on ice and load 40ul onto the array. 6. Assemble Hybridization Chamber 7 Hybridize at 65°C for 17 hours.
|
| Scan protocol |
Scan region is set to Scan Area (61 × 21.6 mm).Scan resolution (μm) is set to 5um. Dye channel is set to Red & Green. Green PMT is set to 100% (high); 10% (low) Red PMT is set to 100 % ( high); 10 % ( low). Check eXtended Dynamic Range check box for both High and low PMT scanning 3. Scan.
|
| Description |
W303 cln1,2,3∆ MET3pr-CLN2 cells 5 minutes after methionine addition
|
| Data processing |
Use the Feature Extraction (FE) software v9.5.
|
| |
|
| Submission date |
Jun 07, 2011 |
| Last update date |
Jun 11, 2011 |
| Contact name |
Umut Eser |
| E-mail(s) |
umuteser@stanford.edu
|
| URL |
http://web.me.com/skotheim/Site/People.html
|
| Organization name |
Stanford University
|
| Department |
Biology
|
| Lab |
Skotheim Lab
|
| Street address |
337 Campus Drive
|
| City |
Stanford |
| State/province |
CA |
| ZIP/Postal code |
94305 |
| Country |
USA |
| |
|
| Platform ID |
GPL9825 |
| Series (2) |
| GSE29800 |
Lack of G1 cyclin arrest cell cycle synchronization time-course microarray in Glucose |
| GSE29894 |
Cell cycle and G1 cyclins |
|
