|
| Status |
Public on Sep 16, 2010 |
| Title |
P0_Ovary_CTGF_biological rep2 |
| Sample type |
RNA |
| |
|
| Source name |
P0 Ovary incubated in the presence of 500 ng/ml CTGF for 24 hours
|
| Organism |
Rattus norvegicus |
| Characteristics |
organ culture date batch: 2
genotype: Sprague-Dawley rat
gender: Female
tissue: Rat ovary
developmental stage: Rat ovary at day P0 incubated for 24 more hours
|
| Treatment protocol |
For each ovary sample from which RNA was collected for microarrays, 1-3 ovaries per filter/well were cultured for 24 hours in the absence (controls) or presence (treated) CTGF (human connective tissue growth factor)(500 ng/mL, PeproTech Inc., NJ USA). Two or three ovaries from the same culture well (from different rat pups out of the same litter) and receiving the same treatment were pooled and homogenized together.
|
| Growth protocol |
Zero-day old (< 8 hour old) female Sprague-Dawley rats (Harlan Laboratories, Inc., USA) were euthanized according to Washington State University IACUC approved protocols and the ovaries removed and cultured whole as described previously [Dole, G., et al., 2008].
|
| Extracted molecule |
total RNA |
| Extraction protocol |
RNA was isolated from whole rat ovaries (2-3 ovaries per each sample) after homogenization in 1 ml Trizol™ reagent (Sigma-Aldritch, USA), according to manufacturer’s instructions.
|
| Label |
biotin
|
| Label protocol |
Biotin-labeled ssDNA were prepared according to the standard Affymetrix protocol from at least 300 ng total RNA (GeneChip® Whole Transcript (WT) Sense Target Labeling Assay Manual, 2005-2209, Affymetrix).
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| |
|
| Hybridization protocol |
Following fragmentation, ssDNA were hybridized on Affymetrix Rat Gene 1.0 ST Array according to standard Affymetrix protocol. Chips were washed and stained in the Affymetrix Fluidics Station 4500.
|
| Scan protocol |
GeneChips were scanned using Affymetrix GeneChip® Scanner 3000.
|
| Description |
Gene expression data from rat P0 Ovary incubated in the presence of 500 ng/ml CTGF for 24 hours
|
| Data processing |
The data were analyzed with Partek Genomic Suite 6.5 beta software (Partek Inc., St. Louis, MO) using RMA, GC-content adjusted algorithm background correction, quintile normalization, median polish methods for probesets summarization, and log values of probes signals using base 2. Organ culture date batch effect was removed.
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| |
|
| Submission date |
Jun 02, 2010 |
| Last update date |
Sep 16, 2010 |
| 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 |
GPL6247 |
| Series (1) |
| GSE22096 |
Induction of Ovarian Primordial Follicle Assembly by Connective Tissue Growth Factor CTGF |
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