|
| Status |
Public on Dec 31, 2011 |
| Title |
Exposed to 30% RH for 18hr , biological rep4 |
| Sample type |
RNA |
| |
|
| Source name |
Female adult, exposed to 30% RH for 18hr
|
| Organism |
Anopheles gambiae |
| Characteristics |
tissue: whole body
genotype: G3
desiccation: Exposed to 30% RH for 18hr
|
| Treatment protocol |
Mosquitoes were exposed to 70% (standard) or 30% (desiccated) RH without any access to sugar and water. A total of 4 cages of mosquitoes were set up simultaneously. For each humidity condition, twelve surviving mosquitoes were collected and at 0 hr (right before the desiccation experiment was initiated), 18 and 36 hr post exposure except in 30% RH treatment 36 hr post exposure did not yield any surviving mosquitoes (9 mosquitoes collected only). All samples were preserved in RNAlater® (Ambion Inc.) and in -80 oC freezer until RNA extraction.
|
| Growth protocol |
There were 250 adult (male:female=1:1) for each cage. Adults had access to raisins and cotton balls saturated with dd-water. Mosquito colonies were maintained by blood feeding on anesthetized rabbits very five days.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Trizol extraction of total RNA was performed according to the manufacturer's instructions.
|
| Label |
Biotin
|
| Label protocol |
3.75 μg of the resulting amplified cDNA material for each sample was first fragmented and subsequently end-labeled using the NuGEN FL-Ovation cDNA Biotin Module V2 (NuGEN Technologies, San Carlos CA)
|
| |
|
| Hybridization protocol |
2.2 μg of the resulting fragmented, biotin-tagged cDNA was placed into a hybridization cocktail (220 μL), with 200 μL actually hybridized at 45°C with rotation for 18 hours (Affymetrix GeneChip Hybridization Oven 640) to probe sets present on an Affymetrix array (GeneChip® Plasmodium/Anopheles Genome Array). The GeneChip arrays were washed and then stained (SAPE, streptavidin-phycoerythrin) on an Affymetrix Fluidics Station 450 using the FS450_0004 fluidics protocol
|
| Scan protocol |
Scanning on an Affymetrix GeneChip 3000 Scanner 7G
|
| Description |
Exposed to 30% RH for 18hr
|
| Data processing |
Results were quantified and analyzed using Expression Console ver.1.1 software (Affymetrix, Inc.) using the PLIER Algorithm default values (Quantification Scale: Linear; Quantification Type: Signal and Detection P-Value; Background: PM-GCBG; Normalization Method: Sketch-Quantile).
|
| |
|
| Submission date |
Nov 17, 2010 |
| Last update date |
Dec 31, 2011 |
| Contact name |
Mei-Hui Wang |
| E-mail(s) |
meihuiw@uci.edu
|
| Phone |
949-824-0249
|
| Fax |
949-824-0249
|
| Organization name |
UC irvine
|
| Street address |
3501B Hewitt Hall UCI
|
| City |
Irvine |
| State/province |
CA |
| ZIP/Postal code |
92697 |
| Country |
USA |
| |
|
| Platform ID |
GPL1321 |
| Series (1) |
| GSE25433 |
Genome-wide Transcriptional Analysis of Genes Associated with Desiccation Stress in Anopheles gambiae |
|
