|
| Status |
Public on Jan 01, 2010 |
| Title |
daf-2 mutants treated with daf-16 RNAi vs. daf-2 mutants treated with empty vector RNAi replicate 3 |
| Sample type |
RNA |
| |
|
| Channel 1 |
| Source name |
daf-2 mutant animals treated with daf-16 RNAi
|
| Organism |
Caenorhabditis elegans |
| Characteristics |
strain: daf-2(e1370)
developmental stage: adult
gender: hermaphrodite
|
| Treatment protocol |
daf-2 mutant animals treated with daf-16 RNAi feeding
|
| Growth protocol |
Culture on daf-16 or empty vector RNAi-expressing bacteria-seeded agar plates (20 centidegree)
|
| Extracted molecule |
polyA RNA |
| Extraction protocol |
Total RNA extracted using Trizol following manufacturer's instructions (Invitrogen)
|
| Label |
Cy3
|
| Label protocol |
5 µg of poly A RNA was reversed transcribed at 42°C for 2 hours in the presence of 200 U SuperScript II Reverse Transcriptase (Invitrogen), random hexamer primers and each dATP, dTTP, dGTP, dCTP, with aa-dUTP. The cDNA was then cleaned up, labeled with Cy3 or Cy5, and quenched and cleaned up again.
|
| |
|
| Channel 2 |
| Source name |
daf-2 mutant animals treated with empty vector RNAi
|
| Organism |
Caenorhabditis elegans |
| Characteristics |
strain: daf-2(e1370)
development: adult
gender: hermaphrodite
|
| Treatment protocol |
daf-2 mutant animals treated with empty vector RNAi
|
| Growth protocol |
Culture on daf-16 or empty vector RNAi-expressing bacteria-seeded agar plates (20 centidegree)
|
| Extracted molecule |
polyA RNA |
| Extraction protocol |
Total RNA extracted using Trizol following manufacturer's instructions (Invitrogen)
|
| Label |
Cy5
|
| Label protocol |
5 µg of poly A RNA was reversed transcribed at 42°C for 2 hours in the presence of 200 U SuperScript II Reverse Transcriptase (Invitrogen), random hexamer primers and each dATP, dTTP, dGTP, dCTP, with aa-dUTP. The cDNA was then cleaned up, labeled with Cy3 or Cy5, and quenched and cleaned up again.
|
| |
|
| |
| Hybridization protocol |
Oligoarray control targets and hybridization buffer (3XSSC, 20mM HEPES pH 7.0, 0.2% SDS) were added, and samples were applied to microarrays enclosed in hybridization chambers. After hybridization for at least 6 hours at 63 centidegree, slides were washed sequentially.
|
| Scan protocol |
Scanned on an Axon Gene Pix scanner.
Images were quantified using Gene Pix Pro version 4.4 (Molecular Devices)
|
| Description |
Biological replicate 3 of 8. daf-2 mutants treated with daf-16 RNAi vs. daf-2 mutants treated with empty vector RNAi
|
| Data processing |
Data were normalized using Acuity version 4.0 (Molecular Devices)
|
| |
|
| Submission date |
Oct 14, 2009 |
| Last update date |
Oct 16, 2009 |
| Contact name |
SEUNG-JAE LEE |
| E-mail(s) |
SEUNGJAELEE1@GMAIL.COM
|
| Phone |
82-54-279-2351
|
| Organization name |
POHANG UNIVERSITY OF SCIENCE AND TECHNOLOGY
|
| Department |
LIFE SCEICNE
|
| Lab |
MOLECULAR GENETICS OF AGING
|
| Street address |
SAN 31 HYOJA-DONG, NAM-GU
|
| City |
POHANG |
| State/province |
GYEONGBUK |
| ZIP/Postal code |
790-784 |
| Country |
South Korea |
| |
|
| Platform ID |
GPL9450 |
| Series (2) |
| GSE18561 |
Adult C. elegans: Control daf-2 mutants treated with daf-16 RNAi vs. daf-2 mutants treated with empty vector RNAi |
| GSE18563 |
Glucose Shortens the Lifespan of Caenorhabditis elegans by Down-Regulating Aquaporin Gene Expression |
|
