|
| Status |
Public on Feb 08, 2013 |
| Title |
dsHSF1-treated male rep1 |
| Sample type |
RNA |
| |
|
| Channel 1 |
| Source name |
whole adult, dsHS1
|
| Organism |
Anopheles gambiae |
| Characteristics |
gender: male
genotype/variation: HSF1 knockdown
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA from 4-d-old dsLacZ-treated controls and HSF-silenced males was extracted using TRIzol reagent (Life Technologies), following protocols set according to manufacturer’s instructions. RNA preparation, labelling, hybridization and data analysis were performed by the Oxford Gene Technology (OGT) company. In particular, RNA was further cleaned up using the RNeasy Mini Kit (Qiagen) followed by ethanol precipitation. Sample passing the purity metrics (260/280 and 260/230 ratio) were considered for labeling and hybridization procedures. Labeling was done with cyanine 3 (dsHSF1 and dsHSF123) and cyanine 5 (control dsLacZ).
|
| Label |
cy3
|
| Label protocol |
Labeling was done with cyanine 3 (dsHSF1 and dsHSF123) and cyanine 5 (control dsLacZ).
|
| |
|
| Channel 2 |
| Source name |
whole adult, dsLacZ
|
| Organism |
Anopheles gambiae |
| Characteristics |
gender: male
genotype/variation: control (dsLacZ)
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA from 4-d-old dsLacZ-treated controls and HSF-silenced males was extracted using TRIzol reagent (Life Technologies), following protocols set according to manufacturer’s instructions. RNA preparation, labelling, hybridization and data analysis were performed by the Oxford Gene Technology (OGT) company. In particular, RNA was further cleaned up using the RNeasy Mini Kit (Qiagen) followed by ethanol precipitation. Sample passing the purity metrics (260/280 and 260/230 ratio) were considered for labeling and hybridization procedures. Labeling was done with cyanine 3 (dsHSF1 and dsHSF123) and cyanine 5 (control dsLacZ).
|
| Label |
cy5
|
| Label protocol |
Labeling was done with cyanine 3 (dsHSF1 and dsHSF123) and cyanine 5 (control dsLacZ).
|
| |
|
| |
| Hybridization protocol |
Oligoarray control targets and hybridization buffer (Agilent In Situ Hybridization Kit Plus) were added, and samples were applied to microarrays enclosed in Agilent SureHyb-enabled hybridization chambers. After hybridization, slides were washed sequentially.
|
| Scan protocol |
Scanned on an Agilent G2565AA scanner. Images were quantified using Agilent Feature Extraction Software (version 9.5.3.1)
|
| Data processing |
Data analysis was performed by OGT according to a standardized procedure, as implemented by the software applications Feature Extraction (version 9.5.3.1.), and Genespring GX (version 11.0). Expression data was first normalised using linear and loess normalization in Feature Extraction, then it was imported into Genespring where it was converted into normalised log ratios (log2 of cy3 divided by cy5) (signal divided by control).
|
| |
|
| Submission date |
Feb 11, 2011 |
| Last update date |
Feb 08, 2013 |
| Contact name |
tania dottorini |
| Organization name |
University of Perugia
|
| Department |
Dept. Exp.Medicine and Bichem. Sci.
|
| Street address |
via del Giochetto
|
| City |
Perugia |
| ZIP/Postal code |
06126 |
| Country |
Italy |
| |
|
| Platform ID |
GPL13157 |
| Series (1) |
| GSE27233 |
Regulation of Anopheles gambiae male accessory gland genes influences post-mating response in female |
|
