|
| Status |
Public on Dec 01, 2018 |
| Title |
DH-43 |
| Sample type |
RNA |
| |
|
| Source name |
Test sample-2_48 hrs 0.312 µM Methyl parathion
|
| Organism |
Drosophila melanogaster |
| Characteristics |
age: 1 day old flies
gender: Male
|
| Treatment protocol |
Drosophila were exposed to 0.0312 & 0.312 µM Methyl parathion concentrations for 24 and 48hrs, along with DMSO(< 0.03%) controls.
|
| Growth protocol |
Drosophila was maintained on standard Drosophila food (maize powder, sugar, yeast, nipagin and propionic acid) at 23-25˚C.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
RNA extraction was perforemd by using Qiagen Rneasy mini kit with Dnase Treatment.
|
| Label |
Cy3
|
| Label protocol |
The samples for Gene expression were labeled using Agilent Quick-Amp labeling Kit (p/n5190-0442). 500ng each of total RNA were reverse transcribed at 40°C using oligo dT primer tagged to a T7 polymerase promoter and converted to double stranded cDNA. Synthesized double stranded cDNA were used as template for cRNA generation. cRNA was generated by in vitro transcription and the dye Cy3 CTP(Agilent) was incorporated during this step. The cDNA synthesis and in vitro transcription steps were carried out at 40°C. Labeled cRNA was cleaned up using Qiagen RNeasy columns (Qiagen, Cat No: 74106) and quality assessed for yields and specific activity using the Nanodrop ND-1000.
|
| |
|
| Hybridization protocol |
600ng of labeled cRNA sample were fragmented at 60ºC and hybridized on to a Agilent designed Drosophila_GXP_8x60K (AMADID: 039596) arrays. Fragmentation of labeled cRNA and hybridization were done using the Gene Expression Hybridization kit of (Agilent Technologies, In situ Hybridization kit, Part Number 5190-0404). Hybridization was carried out in Agilent’s Surehyb Chambers at 65º C for 16 hours.
|
| Scan protocol |
Agilent Microarray Scanner (Agilent Technologies, Part Number G2600D).
|
| Description |
Gene expression after 48hrs 0.312 µM Methyl parathion
|
| Data processing |
Images were quantified using Feature Extraction Software ( Agilent). Feature extracted raw data was analyzed using GeneSpring GX software from Agilent. Normalization of the data was done in GeneSpring GX using the percentile shift normalization method .
|
| |
|
| Submission date |
Apr 25, 2017 |
| Last update date |
Dec 01, 2018 |
| Contact name |
Genotypic technology |
| E-mail(s) |
sudha.rao@genotypic.co.in
|
| Organization name |
Genotypic Technology
|
| Street address |
259, Apoorva 4th cross,80 feet Road,RMV 2ND STAGE
|
| City |
Bangalore |
| State/province |
Karnataka |
| ZIP/Postal code |
560094 |
| Country |
India |
| |
|
| Platform ID |
GPL23370 |
| Series (2) |
| GSE98164 |
Organismal sensitivity to environmental agents induced toxicity: A comparative ecotoxicogenomics approach [Drosophila] |
| GSE98166 |
Organismal sensitivity to environmental agents induced toxicity: A comparative ecotoxicogenomics approach |
|
