|
| Status |
Public on Nov 01, 2012 |
| Title |
Genistein EC20 C |
| Sample type |
RNA |
| |
|
| Source name |
48 hpf embryos
|
| Organism |
Danio rerio |
| Characteristics |
tissue: whole embryo
age: 48 hours post fertilization
treatment: 2.4mg/l genistein
|
| Treatment protocol |
Per concentration and replicate, 24 newly fertilized embryos (<2hpf) were exposed to 2.2 mg/l (EC10) and 2.4 mg/l (EC20) genistein for 48 hours in 96 well plates. After exposure, each group of 24 fish embryos was pooled together, snap-frozen over dry ice and stored at –80°C.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was isolated from frozen, pooled embryos by homogenization in TRIreagent (Sigma-Aldrich, T9424) followed by extraction with chloroform and ethanol and final purification using RNeasy Mini Spin Columns (QIAGEN, Hilden, Germany) according to the manufacturers’ protocols. RNA concentration was measured using a NanoDrop 1000 (Thermo Fisher Scientific, Schwerte, Germany) and RNA quality was verified on a Bioanalyzer 2100 (Agilent Technologies, Santa Clara, USA).
|
| Label |
Cy3
|
| Label protocol |
We carried out cDNA synthesis, cRNA Cy3 labeling, amplification and hybridization according to the manufacturer’s protocols (Quick Amp Labeling Kit; Agilent Technologies), using 500ng RNA as starting material.
|
| |
|
| Hybridization protocol |
Hybridization was carried out according to the Agilent one-color microarray hybridization protocol (One-Color Microarray-Based Gene Expression Analysis, version 5.7, Agilent Technologies)
|
| Scan protocol |
Microarrays were scanned immediately after wahsing with the Agilent Microarray Scanner (Agilent Technologies)
|
| Description |
Gene expression of embryos exposed to 2.4mg/l genistein after 48hpf
|
| Data processing |
Data were extracted and processed using Agilent Feature Extraction Software applying default parameters (protocol GE1-v5_95_Feb07, QC metric set GE1_QCMT_Feb07)
|
| |
|
| Submission date |
Dec 21, 2011 |
| Last update date |
Nov 01, 2012 |
| Contact name |
Viktoria Schiller |
| E-mail(s) |
schiller@molbiotech.rwth-aachen.de
|
| Phone |
0049 (0)241 6085 12272
|
| Organization name |
Fraunhofer IME
|
| Department |
UNIFISH
|
| Street address |
Forckenbeckstrasse 6
|
| City |
Aachen |
| State/province |
NRW |
| ZIP/Postal code |
52074 |
| Country |
Germany |
| |
|
| Platform ID |
GPL6457 |
| Series (1) |
| GSE34616 |
Transcriptome response to genistein in zebrafish embryos |
|
