|
| Status |
Public on Sep 22, 2008 |
| Title |
Ovary_Line FL1_biological rep6_technical rep2 |
| Sample type |
RNA |
| |
|
| Source name |
Sample pools, ovaries from six animals, metestrous stage
|
| Organism |
Mus musculus |
| Characteristics |
mouse line selected for high fertility performance
about six week old females at metestrous stage
|
| Treatment protocol |
Line FL1 was selected for an index trait, combining litter size (LS0) and litter weight (LW0) at birth in primiparous females (Index (I) = 1.6 x LS0 + LW0)) to generation 130
|
| Growth protocol |
Mice were kept in Makrolon-cages of 30 x 12.5 x 12.5 cm with free access to pellet concentrate and water. The light regime was LD 12:12, the room temperature was standardized to 22.5 °C.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was prepared with the RNeasey Mini Kit with simultaneous removal of genomic DNA with RNAse free DNAse (Qiagen)
|
| Label |
Biotin
|
| Label protocol |
Biotinylated cRNA were prepared according to the standard Affymetrix protocol from 10 ug total RNA
|
| |
|
| Hybridization protocol |
The fragmented cRNA was hybridized overnight (45°C) to the Affymetrix GeneChip MOE 430A mouse expression arrays. Each sample pool was hybridized to two arrays to reduce the technical variation. Arrays were then washed using the GeneChip Fluidics Station (Affymetrix) according to the manufacturer’s protocol and stained by R-Phycoerythrin Streptavidin (Molecular Probes). This was followed by an antibody amplification procedure using a biotinylated anti-streptavidin antibody (Vector laboratories) and goat IgG (Sigma).
|
| Scan protocol |
Scanning was carried out with 3µm resolution, 488nm excitation and 570nm emission wavelengths employing the GeneArray Scanner 2500 (Hewlett Packard)
|
| Description |
Ovaries from 30 animals of line FL1 and of the control line DUKsi were collected at the metestrous stage to identify genes that are involved in increasing the ovulation number.
|
| Data processing |
For calculation of signal intensity the Affymetrix Microarray Suite 5.0 Software (MAS5) was used. All arrays were normalized by scaling to reach a common target intensity.
|
| |
|
| Submission date |
Apr 09, 2008 |
| Last update date |
Sep 22, 2008 |
| Contact name |
Jens Vanselow |
| E-mail(s) |
vanselow@fbn-dummerstorf.de
|
| Phone |
+49 38208 68750
|
| Organization name |
Leibniz Institute for Farm Animal Biology
|
| Department |
Reproductive Biology
|
| Lab |
Experimental Reproductive Biology
|
| Street address |
Wilhelm-Stahl-Allee 2
|
| City |
Dummerstorf |
| ZIP/Postal code |
18196 |
| Country |
Germany |
| |
|
| Platform ID |
GPL339 |
| Series (1) |
| GSE11113 |
Expression profiling of a high-fertility mouse line by microarray analysis and qPCR. |
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