|
| Status |
Public on Nov 21, 2023 |
| Title |
Animal cap explants of uninjected embryos biological rep2 |
| Sample type |
RNA |
| |
|
| Source name |
Animal cap explants, uninjected
|
| Organism |
Xenopus laevis |
| Characteristics |
tissue: Animal cap explant
treatment: Uninjected
Stage: dissected at stage 8 and incubated for 3 hours
|
| Treatment protocol |
chordin mRNA (25 pg/cell) was injected into the animal pole of all blastomeres at the four-cell stage. Animal cap explants were prepared at stage 8 and incubated in 1x Steinberg's solution for 3 hours followed by collection for microarray analysis. For uninjected embryos, embryos were cultured and treated in the same way except injection.
|
| Growth protocol |
Xenopus laevis embryos were obtained by in vitro fertilization and cultured in 0.1× MBS at 22°C. Embryos at stage 4 were transferred into 4% Ficoll in 0.1×MBS and cultured overnight at 15°C in 4% Ficoll in 0.1x MBS until stage 8 (mid-blastula).
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was extracted using TRIzol reagent.
|
| Label |
biotin
|
| Label protocol |
Biotinylated cRNA was prepared from 250 ng total RNA using the GeneChip 3’ IVT Express Kit (Affymetrix) according to the manufacturer’s instruction.
|
| |
|
| Hybridization protocol |
Following fragmentation, 15 ug of cRNA was hybridized for 16 hr at 45°C on Affymetrix GeneChip Xenopus laevis Genome 2.0 Arrays. GeneChips were washed and stained in the Affymetrix Fluidics Styation 450.
|
| Scan protocol |
GeneChips were scanned using an Affymetrix GeneChip Scanner.
|
| Description |
Uninjected embryos were cultured until stage 8. Animal cap explants were prepared at stage 8 and incubated for 3 hours followed by collection for microarray analysis.
|
| Data processing |
The CEL files were analyzed with RMA algorithm on GeneSpring X8 with default analysis settings.
|
| |
|
| Submission date |
Jan 03, 2018 |
| Last update date |
Nov 21, 2023 |
| Contact name |
Toshiyasu Suzuki |
| E-mail(s) |
T.Suzuki@beatson.gla.ac.uk
|
| Organization name |
Kyoto University
|
| Department |
Department of Cell and Developmental Biology, Graduate school of Biostudies
|
| Street address |
Sakyo-ku
|
| City |
Kyoto |
| ZIP/Postal code |
606-8502 |
| Country |
Japan |
| |
|
| Platform ID |
GPL10756 |
| Series (2) |
| GSE108684 |
Fam46a is a regulator for gene expression in secretory tissues and plays an important role in development and homeostasis [Chordin] |
| GSE108692 |
Fam46a is a regulator for gene expression in secretory tissues and plays an important role in development and homeostasis |
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