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Status |
Public on Dec 31, 2011 |
Title |
Exposed to 70% RH for 18hr , biological rep1 |
Sample type |
RNA |
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Source name |
Female adult, exposed to 70% RH for 18hr
|
Organism |
Anopheles gambiae |
Characteristics |
tissue: whole body genotype: G3 desiccation: Exposed to 70% 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.
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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)
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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
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Scan protocol |
Scanning on an Affymetrix GeneChip 3000 Scanner 7G
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Description |
Exposed to 70% 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).
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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 |
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