|
| Status |
Public on May 20, 2008 |
| Title |
large_follicle_granulosa_cell_rep2 |
| Sample type |
RNA |
| |
|
| Source name |
granulosa cells of large follicles (>10 mm)
|
| Organism |
Bos taurus |
| Characteristics |
granulosa cells of large follicles (>10 mm) from follicular stage ovaries of young nonpregnant cycling heifers
granulosa cells were isolated by dissection (i.e. scraped and flushed) from the fresh tissue as previously described (Skinner and Osteen, 1988). Cell preparations have been characterized cytochemically to be greater that 98% pure and contain less than 1% contamination with endothelial cells.
|
| Treatment protocol |
Bovine ovaries were obtained from of young nonpregnant cycling heifers less then 10 min after death. Ovaries were delivered fresh on ice to laboratory. Only follicular stage ovaries were used as assessed by morphological criteria that include the absence of a corpus luteum and the presence of developing large antral follicles.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Cells from 20-40 follicles of the same size were pooled to make each RNA sample. RNA was extracted using the Trizol reagent according to manufacturers instructions (Sigma, St. Louis, MO).
|
| Label |
biotin
|
| Label protocol |
standard Affymetrix procedures
|
| |
|
| Hybridization protocol |
standard Affymetrix procedure
|
| Scan protocol |
standard Affymetrix procedure
|
| Description |
RNA from theca cell of large follicles
|
| Data processing |
Auto-scale ON (1-1000). Scaling = all probe sets, Target Signal 125. The microarray chip image data were converted to numerical data with GeneChip Operating Software (Affymetrix) using MAS-5 preprocessing algorithm.
The absolute analysis from MAS was imported into GeneSpring 7.3 software (Silicon Genetics, Redwood City, CA). The developing follicle data were normalized within GeneSpring using the default/recommended normalization methods. These include setting of signal values below 0.01-0.01, total chip normalization to the 50th percentile, and normalization of each gene to the median.
|
| |
|
| Submission date |
May 19, 2008 |
| Last update date |
May 19, 2008 |
| Contact name |
Michael K Skinner |
| E-mail(s) |
skinner@mail.wsu.edu
|
| Organization name |
WSU
|
| Department |
SBS
|
| Street address |
Abelson 507
|
| City |
Pullman |
| State/province |
WA |
| ZIP/Postal code |
99163 |
| Country |
USA |
| |
|
| Platform ID |
GPL2112 |
| Series (1) |
| GSE11495 |
Regulation of granulosa and theca cell transcriptomes during ovarian antral follicle development. |
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