|
| Status |
Public on Dec 07, 2017 |
| Title |
Male of wild-type N2, fasting biological rep3 |
| Sample type |
RNA |
| |
|
| Source name |
Males subjected to 48 hors of fasting, beginning at day 2 of adulthood
|
| Organism |
Caenorhabditis elegans |
| Characteristics |
genetic background: N2
genotype: wild type
tissue: Whole body
age: Day 4 of adulthood
gender: male
treatment: fasted
|
| Treatment protocol |
At the young adult stage, worms were transferred to NGM plats containing 200 ug/ml of 5’fluoro-2’deoxyuridine (FUdR) to prevent progenies from hatching. At day 2 of adulthood, worms were devided into 2 groups, fed and fasting. In the fed group, worms were allowed to feed ad libitum for 2 days. In the fasting group, worms were placed on NGM plates withour food and subjected to 48 hours of fasting.
|
| Growth protocol |
Synchronized wild-type N2 worms were grown on NGM plates at 20 C. At the young adult stage, males were separated from hermaphrodites by filtration through a 37 mm nylon mesh.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Trizol extraction of total RNA was performed according to the manufacturer's instructions with a minor modification (the addition of sterilized glass beads for homogenization). The extracted RNA was purified and treated with DNase using an RNeasy Mini Kit (Qiagen)
|
| Label |
biotin
|
| Label protocol |
Biotinylated cRNA were prepared according to the standard Affymetrix protocol from 250 ng total RNA (GeneChip 3' IVT Express kit mannual, Affymetrix). IVT incubation time was 16 hours.
|
| |
|
| Hybridization protocol |
Following fragmentation, 10 ug of cRNA were hybridized for 16 hr at 45C on GeneChip C. elegans Genome Array. GeneChips were washed and stained in the Affymetrix Fluidics Station 450.
|
| Scan protocol |
GeneChips were scanned using the Affymetrix GeneChip Scanner.
|
| Description |
Gene expression data from adult worms after the completion of development
|
| Data processing |
Images from scanned chips were processed by using the default settings of the AGCC viewer. The Affymetrix output (CEL files) was imported into GeneSpring GX 12.0 (Agilent Technologies) microarray analysis software for the statistical analysis and presentation of the expression profiles. The CEL files were individually scaled to a median target signal of 500. The probe intensities, expression signals and fold changes of all genes (probe sets) were calculated using MAS5 implemented in GeneSpring software.
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| |
|
| Submission date |
Apr 16, 2012 |
| Last update date |
Dec 07, 2017 |
| Contact name |
Eisuke Nishida |
| E-mail(s) |
nishida@lif.kyoto-u.ac.jp
|
| Phone |
+81-75-753-4230
|
| Organization name |
Graduate School of Biostudies, Kyoto University
|
| Department |
Department of Cell and Developmental Biology
|
| Street address |
Kitashirakawa, Sakyo-ku
|
| City |
Kyoto |
| ZIP/Postal code |
606-8502 |
| Country |
Japan |
| |
|
| Platform ID |
GPL200 |
| Series (2) |
| GSE37303 |
Fasting induced changes in gene expression profiles of C. elegans |
| GSE37305 |
Age and Fasting in C. elegans |
|
