|
| Status |
Public on Apr 10, 2007 |
| Title |
Expression in spo11Y135F strain at meiosis 4 hr |
| Sample type |
RNA |
| |
|
| Source name |
4 hrs after transfer into sporulation medium
|
| Organism |
Saccharomyces cerevisiae |
| Characteristics |
strain: RKD1312
background: SK1
genotype: spo11Y135F
|
| Treatment protocol |
Cells were washed twice with water, frozen in liquid nitrogen, and stored at -80C.
|
| Growth protocol |
Cells were grown on a YPD plate for 2 days at 30C. The cells were grown to a density of 4.0x10^7 cells/ml at 30C in SPS presporulation medium (0.5% yeast extract, 1% peptone, 0.17% yeast nitrogen base w/o amino acid and ammonium sulfate, 1% potassium acetate, 0.5% ammonium sulfate, usual level fo amino acids, 50 mM potassium phthalate buffer pH 5.0, and antifoam). The cells were suspended in sporulation medium (1% potasium acetate, 1/5 level of amino acids, and polypropylene glycol) to a density of 2x10^7 cells/ml, and incubated for 4 hrs at 30C for sporulation.
|
| Extracted molecule |
polyA RNA |
| Extraction protocol |
Total RNA was prepared using glass beads and RNeasy Maxi Kit (QIAGEN). mRNA was isolated the total RNA using Oligotex-dT30 (TaKaRa). These processes were performed according to the manufacturer's instructions.
|
| Label |
biotin
|
| Label protocol |
Biotinylated cRNA were prepared according to the standard Affymetrix protocol from 3 microg mRNA (Expression Analysis Technical Manual, 2001, Affymetrix).
|
| |
|
| Hybridization protocol |
Following fragmentation, 15 microg of cRNA were hybridized for 16 hr at 45C on GeneChip YG-S98. GeneChips were washed and stained using EukGE-WS1 ver3 protocol in the Affymetrix Fluidics Station 400.
|
| Scan protocol |
GeneChips were scanned using the Hewlett-Packard GeneArray Scanner.
|
| Description |
Gene expression data from diploid cells in prophase.
|
| Data processing |
The data were analyzed with Microarray Suite version 5.0 (MAS 5.0) using Affymetrix default analysis settings. The comparison with GSM153627 was performed with MAS 5.0 using Affymetrix default analysis settings. See Comparison data table on Series GSE6620 record.
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| |
|
| Submission date |
Dec 29, 2006 |
| Last update date |
Jan 03, 2007 |
| Contact name |
Kazuto Kugou |
| E-mail(s) |
kkugou@kazusa.or.jp
|
| Organization name |
Kazusa DNA Research Institute
|
| Department |
Department of Frontier Research
|
| Lab |
Laboratory of Cell Engineering
|
| Street address |
2-6-7 KazusaKamatatri
|
| City |
Kisarazu |
| State/province |
Chiba |
| ZIP/Postal code |
292-0818 |
| Country |
Japan |
| |
|
| Platform ID |
GPL90 |
| Series (1) |
| GSE6620 |
Expression data from wild type, mre11delta, rad50delta, and spo11Y135F at meiosis 0 hr and 4 hr. |
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