|
| Status |
Public on Dec 29, 2007 |
| Title |
Cont MO |
| Sample type |
RNA |
| |
|
| Source name |
zebrafish embryos at 24 hpf
|
| Organism |
Danio rerio |
| Characteristics |
AB line
|
| Treatment protocol |
For injection, MOs were at a concentration of about 5ng each embryo. After injection, embryos were incubated at 28.5°C in Embryo Medium. The embryos at 24 hpf were washed with PBS and extracted RNA with Trizol
|
| Growth protocol |
A breeding colony of zebrafish (Danio rerio) were maintained at 28.5°C on a 14 h (hour) light/10 h dark cycle
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Trizol extraction of total RNA was performed according to the manufacturer's instructions.
|
| Label |
biotin
|
| Label protocol |
cRNAs were generated and hybridized on Affymetrix DNA chips according to the Affymetrix protocol, total RNA (8 μg) was used to prepare double-stranded cDNA with the one-cycle cDNA synthesis kit (Affymetrix, USA). The cDNA was then used as a template to synthesize a biotinylated cRNA (16 hr, 37°C) with the IVT kit (Affymetrix, USA). In vitro transcription products were purified with the IVT cleanup module and approximately 35 μg of cRNA were treated with the fragmentation buffer (35' at 94°C).
|
| |
|
| Hybridization protocol |
Following fragmentation, 10 ug of cRNA were hybridized for 16 hr at 37°C on Affymetrix GeneChip Zebrafish Genome Array. GeneChips were washed and stained in the Affymetrix Fluidics Station 450.
|
| Scan protocol |
GeneChips were scanned using the GeneArrayTM scanner 3000.
|
| Description |
Gene expression data from embryos at 24 hpf injected with control Morpholino (five mismatch DrC1q-like MO)
|
| Data processing |
The data were analyzed with Microarray Suite version 5.0 (MAS 5.0) using Affymetrix default analysis settings and global scaling as normalization method. The trimmed mean target intensity of each array was arbitrarily set to 500.
|
| |
|
| Submission date |
Aug 16, 2007 |
| Last update date |
Aug 14, 2011 |
| Contact name |
Jie Mei |
| E-mail(s) |
kevinmei@ihb.ac.cn
|
| Organization name |
Institute of Hydrobiology
|
| Department |
Chinese Academy of Sciences
|
| Lab |
Wuhan Center for Developmental Biology
|
| Street address |
7# Donghu South Road
|
| City |
Wuhan |
| State/province |
Hubei |
| ZIP/Postal code |
430072 |
| Country |
China |
| |
|
| Platform ID |
GPL1319 |
| Series (1) |
| GSE8800 |
Expression data from embryos injected with DrC1q-like morpholino and control morpholino |
|
