|
| Status |
Public on Feb 05, 2013 |
| Title |
ERR1/ERR2 mutant larvae at late-L3, normoxia biological rep3 |
| Sample type |
RNA |
| |
|
| Source name |
Drosophila larvae at partial clear gut stage (-10 to -4 hr prior to pupariation) grown at 25C on yeast paste in nomoxia
|
| Organism |
Drosophila melanogaster |
| Characteristics |
tissue: whole larvae
genotype: dERR1/dERR2 (product of the cross of parents dERR1/TM3,tw-GFP x dERR2/TM3,tw-GFP)
age: late-L3 partial clear gut (-10 to -4 hrs prior to metamorphic onset) as judged by blue food content in gut
|
| Treatment protocol |
Appropriately staged animals were moved to fresh agar plates ~36 hours after hatching and allowed to continue development. Wandering larvae were manually selected for size, vitality, and appropriate spiracle formation, and, importantly, blue food content of the gut. Selected animals were allowed to age an additional 6 hours at 25ºC (normoxia). Alternatively, animals were placed in an airtight Modular Incubator Chamber for 6 hours at 25ºC after a gas mixture containing 4% oxygen balanced with nitrogen was flashed into the chamber (hypoxic treatment). After either treatment, larvae were rinsed with 1xPBS and frozen at -80C until further processing.
|
| Growth protocol |
Fly eggs were collected onto egg caps with yeast paste . The caps were replaced after overnight incubation periods and kept at 25ºC, until mid-L2. At mid-L2, larvae were transferred to a fresh egg cap with blue yeast paste (0.3% bromophenol blue), and allowed to develop until achieving the partial clear gut L3 stage (-10 to -4 hrs prior to metamorphic onset). Appropriately staged animals were moved to fresh agar plates and allowed to age an additional 6 hours at 25ºC.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Larvae were placed in TRIzol (Invitrogen, Carlsbad, CA) and homogenized using VWR disposable pellet mixer. Total RNA was isolated using a TRIzol/RQ1 DNase (Promega, Madison, WI) hybrid extraction protocol.
|
| Label |
biotin
|
| Label protocol |
Template labelings were performed using the GeneChip 3' IVT Express Kit according to the manufacturer’s specifications (Affymetrix, Santa Clara, CA).
|
| |
|
| Hybridization protocol |
Hybridizations to Affymetrix GeneChip Drosophila Genome 2.0 arrays were performed using the manufacturers recommendations.
|
| Scan protocol |
Every chip was scanned at a high resolution by the Affymetrix GeneChip® Scanner 3000 according to the GeneChip Expression Analysis Technical Manual procedures
|
| Description |
Gene expression data from late-L3 partial clear gut dERR mutant animals hand sorted and treated in normoxia
|
| Data processing |
Raw data were normalized using RMA
|
| |
|
| Submission date |
Oct 15, 2012 |
| Last update date |
Feb 05, 2013 |
| Contact name |
Keith D Baker |
| E-mail(s) |
kdbaker@vcu.edu
|
| Phone |
8046284498
|
| Organization name |
VCU School of Medicine
|
| Department |
Biochemistry & Molecular Biology
|
| Street address |
1220 E. Broad St.
|
| City |
Richmond |
| State/province |
VA |
| ZIP/Postal code |
23298 |
| Country |
USA |
| |
|
| Platform ID |
GPL1322 |
| Series (1) |
| GSE33100 |
HIF- and non-HIF-Regulated Hypoxic Responses Require the Estrogen-Related Receptor in Drosophila |
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