|
| Status |
Public on Mar 31, 2010 |
| Title |
Mercuric chloride treated for 48hr, 2 |
| Sample type |
RNA |
| |
|
| Channel 1 |
| Source name |
Zebrafish liver tissue
|
| Organism |
Danio rerio |
| Characteristics |
tissue type: liver
agent: Mercuric chloride
time point: 48hr
|
| Treatment protocol |
Treatment for microarray analyses was carried out in 4 days, using one concentration (200 µg/L) of mercuric chloride with liver RNA sampling at 8, 24, 48, and 96 hours, respectively. Treatment was conducted using triplicate groups of four zebrafishes.
|
| Growth protocol |
Zebrafish were reared at room temperature (27°C)
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA of tissue samples were extracted using Trizol reagent (Invitrogen) according to the manufacturers instructions.
|
| Label |
Cy5
|
| Label protocol |
For fluorescence labeling of cDNAs, 20 µg of total RNA from the reference and experimental samples were reverse transcribed in the presence of Cy3-dUTP and Cy5-dUTP (Amersham Inc.), respectively. Labeled cDNA were pooled, concentrated, and resuspended in DIG EasyHyb (Roche Applied Science) buffer for hybridization.
|
| |
|
| Channel 2 |
| Source name |
Zebrafish liver tissue
|
| Organism |
Danio rerio |
| Characteristics |
reference: Reference RNA was obtained by pooling equal amount of male and female total RNA extracted from liver tissues of wild-type zebrafish.
|
| Growth protocol |
Zebrafish were reared at room temperature (27°C)
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA of tissue samples were extracted using Trizol reagent (Invitrogen) according to the manufacturers instructions.
|
| Label |
Cy3
|
| Label protocol |
For fluorescence labeling of cDNAs, 20 µg of total RNA from the reference and experimental samples were reverse transcribed in the presence of Cy3-dUTP and Cy5-dUTP (Amersham Inc.), respectively. Labeled cDNA were pooled, concentrated, and resuspended in DIG EasyHyb (Roche Applied Science) buffer for hybridization.
|
| |
|
| |
| Hybridization protocol |
Reference RNA was obtained by pooling equal amount of total RNA extracted from male and female wildtype fish. Reference RNA is co-hybridized with RNA samples extracted either from treated or control fish on a glass array spotted with 16,416 zebrafish oligo probes (16,244 LEADS clusters and 172 copies of β-actin controls)
|
| Scan protocol |
The arrays were scanned using the GenePix 4000B microarray scanner (Axon Instruments, USA) and the generated images with their fluorescence signal intensities were analyzed using GenePix Pro 4.0 image analysis software (Axon Instruments, USA).
|
| Description |
Biological replicate 2 of 3: mercury-treated at 48 Hr
|
| Data processing |
The data were global median normalized.Normalized ratio of means defined by CH1/CH2
|
| |
|
| Submission date |
Nov 03, 2009 |
| Last update date |
Jan 26, 2010 |
| Contact name |
Choong Yong UNG |
| Organization name |
National University of Singapore
|
| Department |
Biological Sciences Department
|
| Lab |
Gong Zhiyuan
|
| Street address |
Science Drive 4
|
| City |
Singapore |
| State/province |
Singapore |
| ZIP/Postal code |
117543 |
| Country |
Singapore |
| |
|
| Platform ID |
GPL2715 |
| Series (1) |
| GSE18861 |
Mercury-Induced Hepatotoxicity in Zebrafish |
|
