|
| Status |
Public on Jul 10, 2019 |
| Title |
Single blastocyst in SAGE 1step 14b |
| Sample type |
RNA |
| |
|
| Source name |
preimplantation mouse embryo
|
| Organism |
Mus musculus |
| Characteristics |
genetic background: B6C3F1 x CD1
tissue: Single blastocyst
age: Theiler stage 5; 4 dpc (96h IVC)
medium: SAGE 1step
cell number: approx 40-50 cells per blastocyst
|
| Treatment protocol |
At the 2-cell stage embryos were split, cultured individually to blastocyst stage, and these blastocysts were lysed in Zymo buffer.
|
| Growth protocol |
Zygotes from B6C3F1 x CD1 mating (stimulated cycle, PMSG + hCG given 48 hours apart) were collected from the oviducts and cultured to the 2-cell stage in KSOM(aa) medium using a 4-well plate under 5% CO2 in air at 37 degrees C. At the 2-cell stage the sister blastomeres were separated from each other and cultured individually in ART medium (GM501 CULT or SAGE 1-Step) or in KSOM(aa) medium in a 96-well plate for additional 72 hours (approx. 96 hours in total).
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Individual blastocysts were lysed in Zymo buffer (Quick RNA micro prep) and total RNA was eluted in water according to the manufacturer's instructions.
|
| Label |
biotin
|
| Label protocol |
Biotinylated cDNA was prepared using GeneChip WT Pico kit, reverse-transcribing 20 micrograms cRNA and, of these, subjecting 5.5 micrograms to labeling.
|
| |
|
| Hybridization protocol |
The GeneChip Hybridization, Wash and Stain Kit was used, for 16 h (+/- 1h) at 45°C with 60 rpm. GeneChips were washed and stained in the Affymetrix Fluidics Station 450.
|
| Scan protocol |
The GeneChips were scanned using the Affymetrix 3000 7G scanner.
|
| Description |
14b
Half embryo in SAGE 1step
|
| Data processing |
The robust multiarray averaging method was applied for background correction, normalization, and probe summarization using the Affymetrix Expression Console.
|
| |
|
| Submission date |
Oct 05, 2018 |
| Last update date |
Jul 10, 2019 |
| Contact name |
Michele Boiani |
| E-mail(s) |
[email protected]
|
| Organization name |
Max Planck Institute Muenster
|
| Street address |
Roentgenstrasse 20
|
| City |
Muenster |
| ZIP/Postal code |
48149 |
| Country |
Germany |
| |
|
| Platform ID |
GPL20258 |
| Series (1) |
| GSE120905 |
Adaptive molecular responses of mouse half-embryos grown in different culture media |
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