|
| Status |
Public on Nov 07, 2012 |
| Title |
Blastocyst 2 (Mouse) |
| Sample type |
RNA |
| |
|
| Source name |
Blastocyst
|
| Organism |
Mus musculus |
| Characteristics |
tissue: Blastocyst
age: 3 to 5 weeks old
strain: B6D2F1
gender: females
|
| Treatment protocol |
Immunosurgery for inner cell mass isolation: Zona pellucida was removed by shortly incubating the blastocysts with Tyrode’s solution (Sigma). Consequently zona-free blastocysts were washed several times with M2 medium (Sigma) and immunosurgery was performed, by incubating the embryos 30 min with rabbit anti-mouse serum (Sigma). Afterwards, embryos were washed several times with DMEM medium (GIBCO) supplemented with 10% of fetal bovine serum. The embryos were then incubated (37°C, 5% CO2) with guinea pig complement serum (Sigma) for another 30 min. Rabbit anti-mouse serum was used 1:30 diluted in DMEM medium; guinea pig complement serum was diluted 1:3 in DMEM.
|
| Growth protocol |
3 to 5 weeks old B6D2F1 females were superovulated with 5IU pregnant mare’s serum gonadotropin (PMSG) (FolligonH Intervet) and 48 h later, with 5IU human chronicle gonadotropin (hCG) (ChorulonH Intervet). Superovulated females were mated with B6D2F1 males for 24 h and then housed separately to the males. Upon isolation morula and blastocyst stage embryos were either stored at -80°C for further RNA extraction or immunosurgery was performed for the isolation of the inner cell masses (ICMs).
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was extracted by using RNeasy Micro Kit (QIAGEN) according to the manufacture’s recommended protocols. All RNA samples were kept at -80°C till they were processed for microarray hybridization.
|
| Label |
biotin
|
| Label protocol |
Biotinylated cRNA were prepared according to the standard Affymetrix protocol from 6 ug total RNA (Expression Analysis Technical Manual, 2001, Affymetrix).
|
| |
|
| Hybridization protocol |
Following fragmentation, 10 ug of cRNA were hybridized for 16 hr at 45C on GeneChip Drosophila Genome Array. GeneChips were washed and stained in the Affymetrix Fluidics Station 400.
|
| Scan protocol |
GeneChips were scanned using the Hewlett-Packard GeneArray Scanner G2500A.
|
| Description |
Blastocyst
|
| Data processing |
RMA summaries using Brainarray CDFs Version 13.0.0 ENSG
|
| |
|
| Submission date |
Nov 06, 2012 |
| Last update date |
Nov 07, 2012 |
| Contact name |
Michal J Okoniewski |
| E-mail(s) |
michal.okoniewski@gmail.com
|
| Organization name |
ETH Zurich
|
| Lab |
Scientific IT
|
| Street address |
Binzmühlestrasse 130
|
| City |
Zurich |
| State/province |
Zurich |
| ZIP/Postal code |
8092 |
| Country |
Switzerland |
| |
|
| Platform ID |
GPL14657 |
| Series (1) |
| GSE42081 |
Cross-Species Genome Wide Expression Analysis during Pluripotent Cell Determination in Mouse and Rat Preimplantation Embryos |
|
