|
Status |
Public on Sep 09, 2013 |
Title |
CH3HgCl_2uM_rep1 |
Sample type |
RNA |
|
|
Source name |
2 uM CH3HgCl
|
Organism |
Caenorhabditis elegans |
Characteristics |
developmental stage: Mixed-stage exposure level: low-toxic exposure time: 24hr
|
Treatment protocol |
Fifty mL aliquots of the mixed-stage C. elegans population were treated to sub- (2 µM HgCl2, 0.75 µM MeHgCl), low- (7.5 µM HgCl2, 2 µM MeHgCl) or high- (20 µM HgCl2, 7.5 µM MeHgCl) toxic mercurial concentrations for 24 hours in a 200 rpm shaker at 20°C.
|
Growth protocol |
Forty L4 stage N2 nematodes were placed on K-agar plates and allowed to grow for 4 days at 20°C. All nematodes were then transferred into a flask containing complete S-medium with OP50 and were grown in a 200 rpm shaker for 4 days at 20°C.
|
Extracted molecule |
total RNA |
Extraction protocol |
Nematodes were then rinsed in 0.1M NaCl and living nematodes were isolated by sucrose floatation, frozen in liquid nitrogen and stored at -80°C. Nematodes were ground with a liquid nitrogen-cooled mortar and pestle, and total RNA was isolated using the RNeasy Midi Kit (Qiagen, Valencia, CA) according to manufacturer’s instructions.
|
Label |
Cy3
|
Label protocol |
Starting with 500ng of total RNA, Cy3 labeled cRNA was produced according to manufacturer’s protocol.
|
|
|
Hybridization protocol |
For each sample, 1.65ug of Cy3 labeled cRNAs were fragmented and hybridized for 17 hours in a rotating hybridization oven.
|
Scan protocol |
Slides were washed and then scanned with an Agilent Scanner.
|
Description |
Gene expression at 24hr after 2uM CH3HgCl treatment US22502532_251506110333_S01_H.tif US22502532_251506110333_S01_L.tif
|
Data processing |
Slides were washed and then scanned with an Agilent Scanner. Data was obtained using the Agilent Feature Extraction software (v9.5), using the 1-color defaults for all parameters.
|
|
|
Submission date |
Apr 28, 2011 |
Last update date |
Sep 11, 2013 |
Contact name |
NIEHS Microarray Core |
E-mail(s) |
microarray@niehs.nih.gov, liuliw@niehs.nih.gov
|
Organization name |
NIEHS
|
Department |
DIR
|
Lab |
Microarray Core
|
Street address |
111 T.W. Alexander Drive
|
City |
RTP |
State/province |
NC |
ZIP/Postal code |
27709 |
Country |
USA |
|
|
Platform ID |
GPL7727 |
Series (1) |
GSE28915 |
Toxicogenomic Responses to Inorganic and Organic Mercury in Caenorhabditis elegans |
|