|
| Status |
Public on Sep 28, 2016 |
| Title |
DominantFollicle_theca-cells_1 |
| Sample type |
RNA |
| |
|
| Source name |
bovine ovarian dominant follicle, theca interna cells
|
| Organism |
Bos taurus |
| Characteristics |
Stage: follicular
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Homogenized in Tri-Reagent (Sigma-Aldrich) for total RNA extraction according to the manufacturer's instructions.
|
| Label |
biotin
|
| Label protocol |
Biotinylated cRNA were prepared according to the standard Affymetrix protocol from 0.3 ug total RNA (Expression Analysis Technical Manual, 2001, Affymetrix).
|
| |
|
| Hybridization protocol |
Following fragmentation, cRNA were hybridized for 16 hr at 42C on Bovine GeneChip Gene 1.0 ST Array. GeneChips were washed and stained in the Affymetrix Fluidics Station 450.
|
| Scan protocol |
GeneChips were scanned using the Affymetrix GeneChip Scanner 3000 7G using the Affymetrix GeneChip Command Console Software
|
| Description |
Isolated from estrogen-active dominant follicles in ovaries of beef cows (75% Red Angus, 25% MARC III) from the physiology herd located at the University of Nebraska Agricultural Research and Development Center. The University of Nebraska-Lincoln Institutional Animal Care and Use Committee approved all procedures and facilities used in this experiment. Estrous cycles of cows were synchronized with a modified Co-Synch protocol using gonadotropin releasing hormone (GnRH) and a controlled internal drug release device (CIDR; 1.38 g progesterone, Zoetis) for 7 days with a PGF2α (25 mg/mL; Lutalyse, Pfizer Animal Health) injection at CIDR removal (Summers et al., 2014). Ovariectomy was performed approximately 36 hours after CIDR removal (Youngquist et al., 1995). Upon ovariectomy, each dominant antral follicle was aspirated/dissected and the granulosa cells (≥ 94% purity), theca cells (≥ 82% purity), and follicular fluid were isolated as described previously (Summers et al., 2014).
|
| Data processing |
Affymetrix Expression Console software was used to perform GC-RMA normalization and export the linear expression data.
|
| |
|
| Submission date |
Jun 20, 2016 |
| Last update date |
Sep 29, 2016 |
| Contact name |
Sarah Romereim |
| E-mail(s) |
sarah.romereim@gmail.com
|
| Organization name |
University of Nebraska-Lincoln
|
| Department |
Animal Science
|
| Lab |
Cupp lab
|
| Street address |
3940 Fair Street
|
| City |
Lincoln |
| State/province |
NE |
| ZIP/Postal code |
68583-0908 |
| Country |
USA |
| |
|
| Platform ID |
GPL16500 |
| Series (1) |
| GSE83524 |
RNA Expression Data from Four Isolated Bovine Ovarian Somatic Cell Types |
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