|
| Status |
Public on Jan 01, 2010 |
| Title |
Male; 0d, rep1 |
| Sample type |
RNA |
| |
|
| Source name |
Pool 5 males (head & thorax only); 0d age
|
| Organism |
Anopheles gambiae |
| Characteristics |
sex: Male
age: 0
|
| Growth protocol |
Anopheles gambiae (strain G3) was reared under standard insectary conditions. Adults were given 10% sucrose solution ad libitum, however no blood meals were provided. Adults emerging within a 24h period were considered a single age class and each age class allocated to separate cages. Adult male and female mosquitoes were collected at 0 (≤ 24 h), 5, 10, 15, 20, 25 and 30 days post-eclosion. The abdomen was dissected and discarded. The heads and thoraces were stored pooled from 5 indivduals.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA extracted using Trizol following manufacturer's instructions
|
| Label |
Cy3
|
| Label protocol |
All arrays were experiments were conducted acording to Agilent Technologies' One-colour Microarray-based Gene Expression Analysis protocol (version 5.7). 1 µg of total RNA and 5µl of 1:5000 dilution Agilent One-colour Spike-in mix were amplified and labelled using the Quick Amp Kit protocols (Agilent Technologies) following manufacturer's protocols.
|
| |
|
| Hybridization protocol |
Labelled cRNA was then purified and quantified before hybridization to the arrays using Agilent Technologies’ Gene Expression Hybridization Kit, SureHyb chambers, Microarray Hybridization oven and standard protocols. Hybridized arrays were washed with Agilent Gene Expression Wash Buffers (without the addition of Triton X102) and Stabilization and Drying Solution.
|
| Scan protocol |
Microarrays were scanned and feature extraction undertaken by technicians at Oxford Gene Technology. Scans were at 5 µm resolution using the extended dynamic range feature of the Agilent Technologies’ DNA microarray scanner (G2505B)
|
| Description |
none
|
| Data processing |
Feature Extraction software (version 9.5.3.1; Agilent Technologies) was used to analyze scanned array image files according to Agilent’s One-colour Gene Expression protocol (GE1-v5_95_Feb07).
|
| |
|
| Submission date |
Sep 21, 2009 |
| Last update date |
Sep 21, 2009 |
| Contact name |
Steven Sinkins |
| E-mail(s) |
steven.sinkins@zoo.ox.ac.uk
|
| Phone |
+44 (0) 1865 281047
|
| Fax |
+44 (0) 1865 281545
|
| Organization name |
University of Oxford
|
| Department |
Department of Zoology
|
| Street address |
South Parks Rd
|
| City |
Oxford |
| ZIP/Postal code |
OX1 3PS |
| Country |
United Kingdom |
| |
|
| Platform ID |
GPL9246 |
| Series (1) |
| GSE18194 |
Transcriptional profiling across adult Anopheles gambiae age |
|
