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Status |
Public on May 21, 2010 |
Title |
Male_Vir D56 |
Sample type |
RNA |
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Source name |
Drosophila mojavensis (15081-1352.22)
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Organism |
Drosophila mojavensis |
Characteristics |
strain: 15081-1352.22 sex: Male treatment: Virgin treatment: Desiccation
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Treatment protocol |
The virgin treatment consisted of keeping the flies in male only or female only vials for eight days transferring them into fresh food vials once at day four. The mated treatment was identical to the virgin treatment for the first seven days, at the beginning of the eighth day male and female vials were combined. Flies remained in these mixed-sex vials for only 24 hours at which time they were separated by sex. At the end of the eight-day period half of the flies from the virgin and mated treatments were placed in new food vials and half in a desiccator. The desiccator was composed of a 30 cm × 30 cm × 30 cm clear acrylic box. Ambient air was pumped through two columns filled with 500 cm3 of Drierite desiccant into the chamber at a rate of 1.5 L per minute. Additionally we placed inside the chamber a container filled with 200 cm3 of Drierite desiccant. Empty 8-dram vials with either six males or females were capped using a cotton ball and placed inside the desiccator. Relative humidity inside the chamber was sampled every minute using a HOBO relative humidity data logger (ONSET Computer Corporation). The relative humidity throughout the experiment was maintained at less than 2%. Prior to commencing the experiment the air in the chamber (with vials) was flushed for 15 minutes. All flies were left inside the desiccator for 40 hours, at which point about 30% of the flies have died (LT30). After this drying period, dead flies were aspirated and discarded while live flies were placed in groups of 15 in 1.5 mL tubes, snap frozen in liquid nitrogen and placed in a -80°C freezer until processing.
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Growth protocol |
Flies were reared in opuntia/banana food in bottle at low density. Adult flies were aged for 7 days in opuntia/banana vials (transferring once on day 4) prior to any treatmet.
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Extracted molecule |
total RNA |
Extraction protocol |
Total RNA extraction was performed using Invitrogen’s PureLink Micro-to-Midi Total Purification System. The suggested manufacturer’s instructions were followed, which included a DNase I digestion to remove any possible genomic DNA contamination. The quality and concentration of the RNA was determined using a NanoDrop spectrophotometer (NanoDrop Technologies).
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Label |
Cy3
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Label protocol |
Labeling was performed by NimbleGen Systems Inc., Madison, WI USA, following their standard operating protocol. See www.nimblegen.com.
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Hybridization protocol |
Hybridization was performed by NimbleGen Systems Inc., Madison, WI, USA following their standard operating protocol. See www.nimblegen.com.
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Scan protocol |
Scanning was performed by NimbleGen Systems Inc., Madison, WI USA, following their standard operating protocol. See www.nimblegen.com.
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Description |
This sample is Male Virgin Desiccation treatment, replicate 2
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Data processing |
The raw data (.pair file) was subjected to RMA (Robust Multi-Array Analysis; Irizarry et al. Biostatistics 4(2):249), quantile normalization (Bolstad et al. Bioinformatics 19(2):185), and background correction as implemented in the NimbleScan software package, version 2.4.27 (Roche NimbleGen, Inc.).
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Submission date |
May 26, 2009 |
Last update date |
May 27, 2009 |
Contact name |
Luciano Matias Matzkin |
E-mail(s) |
lmm0015@uah.edu
|
Phone |
256-824-4326
|
Organization name |
University of Alabama in Huntsville
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Department |
Biological Sciences
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Street address |
301 Sparkman Drive
|
City |
Huntsville |
State/province |
Alabama |
ZIP/Postal code |
35899 |
Country |
USA |
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Platform ID |
GPL8596 |
Series (1) |
GSE16234 |
Transcriptional regulation of metabolism associated with desiccation resistance of the cactophilic Drosophila mojavensis |
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