|
| Status |
Public on Nov 26, 2016 |
| Title |
WT vs VgHop Fat body 2 |
| Sample type |
RNA |
| |
|
| Channel 1 |
| Source name |
Fat body 24 hr after bloodmeal
|
| Organism |
Aedes aegypti |
| Characteristics |
genotype/variation: WT (ORLxRock)
tissue: Fat body
|
| Treatment protocol |
Mosquitoes were provided a blood meal. 24 hours after feeding, mosquitoes were aneshtetized on ice and midguts or fat bodies were collected.
|
| Growth protocol |
Mosquitoes were reared at 27°C in 70% humidity, and adults were maintained on a 10% sucrose solution on a 12-h light/dark cycle, according to standard rearing conditions.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Pools of at least 10 midguts or fat bodies were collected from each treatment type. RNA was extracted from dissected tissues using an RNeasy kit (Qiagen) according to standard protocol.
|
| Label |
Cy3
|
| Label protocol |
Cy5- and Cy3-labeled probes were synthesized from 200 ng of total RNA of each sample using Low Input Quick Amp Labeling Kit from Agilent Technologies
|
| |
|
| Channel 2 |
| Source name |
Fat body 24 hr after bloodmeal
|
| Organism |
Aedes aegypti |
| Characteristics |
genotype/variation: VgHop
tissue: Fat body
|
| Treatment protocol |
Mosquitoes were provided a blood meal. 24 hours after feeding, mosquitoes were aneshtetized on ice and midguts or fat bodies were collected.
|
| Growth protocol |
Mosquitoes were reared at 27°C in 70% humidity, and adults were maintained on a 10% sucrose solution on a 12-h light/dark cycle, according to standard rearing conditions.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Pools of at least 10 midguts or fat bodies were collected from each treatment type. RNA was extracted from dissected tissues using an RNeasy kit (Qiagen) according to standard protocol.
|
| Label |
Cy5
|
| Label protocol |
Cy5- and Cy3-labeled probes were synthesized from 200 ng of total RNA of each sample using Low Input Quick Amp Labeling Kit from Agilent Technologies
|
| |
|
| |
| Hybridization protocol |
Probes prepared from the RNA of transgenic mosquitoes were co-hybridized with probes prepared from the RNA obtained from wild-type mosquitoes. The hybridizations were conducted on an Agilent-based microarray platform using custom-designed whole-genome 8 x 60K A. aegypti microarrays
|
| Scan protocol |
The arrays were scanned with an Agilent High Resolution Scanner (Agilent Technologies Inc., Santa Clara, CA, USA).
|
| Data processing |
Spot intensity was quantified using Agilent Feature Extraction Software. Background-subtracted median fluorescent values were normalized with the LOWESS normalization method, and Cy5/Cy3 ratios from replicate assays were subjected to t-tests at a significance level of p<0.05 using TIGR MIDAS and MeV software. Expression data from replicate assays were averaged with the GEPAS microarray software and logarithm (base 2)-transformed.
|
| |
|
| Submission date |
Nov 25, 2016 |
| Last update date |
Nov 26, 2016 |
| Contact name |
George Dimopoulos |
| E-mail(s) |
gdimopo1@jhu.edu
|
| Organization name |
Johns Hopkins School of Public Health
|
| Department |
Molecular Microbiology and Immunology
|
| Lab |
Dimopoulos Group
|
| Street address |
615 N Wolfe St.
|
| City |
Baltimore |
| State/province |
Maryland |
| ZIP/Postal code |
21205 |
| Country |
USA |
| |
|
| Platform ID |
GPL20714 |
| Series (1) |
| GSE90515 |
Fat body and midgut transcriptome of Wild-type vs. transgenic Aedes aegypti overexpressing JAK/STAT pathway components |
|
