|
| Status |
Public on Dec 21, 2006 |
| Title |
daf19(m86)_daf12(sa204) replicate 2 |
| Sample type |
RNA |
| |
|
| Source name |
daf19(m86)_daf12(sa204)
|
| Organism |
Caenorhabditis elegans |
| Characteristics |
whole worm preparation
embryo
|
| Biomaterial provider |
Michel R. Leroux and Donald G. Moerman
|
| Treatment protocol |
Eggs were prepared from gravid adults using a hypochlorite treatment (www.protocol-online.org), resuspended in 10 mM Tris-EDTA (pH 7.5) and stored at –80 oC. Thawed embryos were disrupted using syringes fit with a 26-gauge needle.
|
| Growth protocol |
solid media
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was isolated using TRIzol reagent (Invitrogen) coupled with phase lock gel tubes (Eppendorf)
|
| Label |
biotin
|
| Label protocol |
Biotinylated cRNA were prepared according to the standard Affymetrix protocol from 5 micrograms total RNA (Expression Analysis Technical Manual, 2004, Affymetrix).
|
| |
|
| Hybridization protocol |
Following fragmentation, 10 micrograms of cRNA were hybridized for 16 hr at 45C on GeneChip C. elegans Genome Array. GeneChip arrays were washed and stained in the Affymetrix Fluidics Station 450 using fluidics script EukGE-WS2v4_450.
|
| Scan protocol |
GeneChip arrays were scanned using the Affymetrix GeneChip Scanner 3000.
|
| Description |
Extracted RNA was subjected to rigorous quality assessment and quantitation using the RNA Nano LabChip Kit (Agilent Technologies) with the 2100 Bioanalyzer (Agilent Technologies). Numerical measures of RNA quality (rRNA ratio, RNA Integrity Number) were employed to ensure the high quality of extracted RNA.
|
| Data processing |
The data were analyzed with GeneChip Operating Software (GCOS) using Affymetrix default analysis settings and scaling to all probe sets with a target signal value of 500 and normalization of 1.
|
| |
|
| Submission date |
Dec 18, 2006 |
| Last update date |
Dec 20, 2006 |
| Contact name |
BCGSC BC Cancer Agency |
| E-mail(s) |
support@mouseatlas.org
|
| Organization name |
Canada's Michael Smith Genome Sciences Centre
|
| Department |
Gene Expression
|
| Lab |
Marco Marra
|
| Street address |
675 West 10th Avenue
|
| City |
Vancouver |
| State/province |
BC |
| ZIP/Postal code |
V5Z 1L3 |
| Country |
Canada |
| |
|
| Platform ID |
GPL200 |
| Series (1) |
| GSE6563 |
Analysis of expression of genes regulated by DAF-19 |
|
