|
| Status |
Public on Jul 01, 2011 |
| Title |
Cbx2_KO_XX_3 |
| Sample type |
RNA |
| |
|
| Source name |
Cbx2_KO_XX gonads at E11.5
|
| Organism |
Mus musculus |
| Characteristics |
genotype/variation: Cbx2_KO_XX
tissue: gonads
age: E11.5
|
| Treatment protocol |
Embryos of the appropriate stage were manually selected under the dissecting scope. Four different genotypes of embryos at E11.5 (XY WT, XX WT, XY Cbx2 KO, and XX Cbx2 KO) were frozen in OCT compound (Sakura Finetechnical, Japan) without fixation. They were sectioned to 30 μm slices and stained with hematoxylin.The developing gonadal cells localized by GATA4 immunostaining and morphology were prepared using a Laser Microdissection System (Leica, Germany). The specimens prepared from two or three individuals were combined into one group, and three groups were prepared for each genotype.
|
| Growth protocol |
Cbx2 KO embryos were generated by heterozygous mice for Cbx2 KO crossing. To stage the embryos accurately, tail somites were counted.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
RNeasy micro kit (QIAGEN) of total RNA was performed according to the manufacturer's instructions.
|
| Label |
biotin
|
| Label protocol |
Total RNAs (15 ng) prepared from these groups were subjected to two-cycle amplification and biotin labeling using MessageAmp™ II aRNA Amplification and MessageAmp™ II-Biotin Enhanced Kit (Ambion, USA), respectively.
|
| |
|
| Hybridization protocol |
Fifteen micrograms of labeled cRNA was fragmented and hybridized to the GeneChip® Mouse Genome 430 2.0 array according to the manufacturer’s instructions (Affymetrix, Santa Clara, CA, USA). GeneChips were washed with R-Phycoerythrin Streptavidin and The GeneChip Hybridization, Wash, and Stain Kit (Affymetrix).
|
| Scan protocol |
GeneChips were scanned using the GeneChip Scanner 3000 (Affymetrix).
|
| Description |
Initiation of sexually dimorphic transcription
Gene expression data from embryonic gonads at E11.5.
|
| Data processing |
Obtained images files were analysed using Affymetrix data suite system, Expression Console (EC) version 1.1. The data were normalized with the RMA (Robust Multichip Average) normalization method in the R statistical package affy. The expression measure is calculated in log base 2 scale.
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| |
|
| Submission date |
May 23, 2011 |
| Last update date |
Jul 01, 2011 |
| Contact name |
Yuko Katoh-Fukui |
| E-mail(s) |
[email protected]
|
| Organization name |
National Inastitute for Longevity Sciences
|
| Street address |
35genngo ,moriokamachi
|
| City |
Oobu |
| ZIP/Postal code |
4748511 |
| Country |
Japan |
| |
|
| Platform ID |
GPL1261 |
| Series (1) |
| GSE29485 |
Expression data from Mouse embryo |
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