|
| Status |
Public on Mar 22, 2011 |
| Title |
Yeast transcriptional response to high (20%) glucose.4 |
| Sample type |
RNA |
| |
|
| Channel 1 |
| Source name |
Yeast cells 1 hour in 2% glucose
|
| Organism |
Saccharomyces cerevisiae |
| Characteristics |
Yeast cells 1 hour in 2% glucose
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Cells were lysed with glass beads in LETS buffer, and total RNA was extracted with phenol and precipitated with 5M LiCl.
|
| Label |
Cy5
|
| Label protocol |
Amino-allyl reverse transcription labeling protocol: Total RNA combined with oligo-dT, 5X First Strand Buffer, 0.1M DTT, 50X aa-dUTP/dNTPs, and SuperScript II.
The cDNA is resuspended and Sodium Bicarbonate is added to sample. Vortex samples and spin down. Add the appropriate dye to the reaction. Vortex samples and spin down. Incubate @ RT - 1 hr in the dark. Clean-up.
|
| |
|
| Channel 2 |
| Source name |
Yeast cells 1 hour in 20% glucose
|
| Organism |
Saccharomyces cerevisiae |
| Characteristics |
Yeast cells 1 hour in 20% glucose
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Cells were lysed with glass beads in LETS buffer, and total RNA was extracted with phenol and precipitated with 5M LiCl.
|
| Label |
Cy3
|
| Label protocol |
Amino-allyl reverse transcription labeling protocol: Total RNA combined with oligo-dT, 5X First Strand Buffer, 0.1M DTT, 50X aa-dUTP/dNTPs, and SuperScript II.The cDNA is resuspended and Sodium Bicarbonate is added to sample. Vortex samples and spin down. Add the appropriate dye to the reaction. Vortex samples and spin down. Incubate @ RT - 1 hr in the dark. Clean-up.
|
| |
|
| |
| Hybridization protocol |
First cDNA-labeled is denaturalised.Place array slide in Hybridization chamber. Add 10ul 3X SSC to slide (well away from spotted array).
Add probe sample onto array area. Apply coverslip PROMPTLY over array. Seal hybridization chamber.
Incubate @ 63°C ~ 16 hrs. by submerging in water bath immediately.
Do different washes with SSC and SDS.
|
| Scan protocol |
GenePix 4000 scanner
|
| Description |
No more description is needed
|
| Data processing |
Intensity obtained in each channel for each pair of microarrays was normalised by Lowess by the Fred Hutchinson Cancer Research Center (FHCRC) Genomics Resource utilizing a pipeline of applications, including GeneTraffic and GTExport (GeneTraffic Export).
|
| |
|
| Submission date |
Nov 20, 2008 |
| Last update date |
Mar 22, 2011 |
| Contact name |
Marcel·lí del Olmo |
| E-mail(s) |
[email protected]
|
| Organization name |
Universitat de València
|
| Street address |
Dr. Moliner, 50
|
| City |
Valencia |
| ZIP/Postal code |
E-46100 |
| Country |
Spain |
| |
|
| Platform ID |
GPL1914 |
| Series (1) |
| GSE13703 |
Transcriptional response of Saccharomyces cerevisiae to high glucose (20%) concentration |
|
