strain: Higgs White Eye age: 2-6 days Sex: female treatment condition: constant dark conditions time point: CT4_DD
Growth protocol
~30 mated non-blood fed female Ae. aegypti Higgs White eye (Wh) strain mosquitoes, aged 2-6 days, were placed in 24 cages, and sequestered in light controlled boxes.
Extracted molecule
total RNA
Extraction protocol
Heads were removed from bodies at 0C on a chilled metal plate and collected. The RNeasy RNA Isolation Kit (Qiagen, Valencia, CA) was used to isolate RNA from each sample, following manufacturer’s instructions. Genomic DNA was instead removed by using the DNA-free Kit (Applied Biosystems, Carlsbad, CA) per manufacturer instructions.
Label
Cy3
Label protocol
Labeling was performed by the Genomics and Bioinformatics Core Facility, University of Notre Dame, using the NimbleGen Single Color Labeling Kit (Roche NimbleGen, Madison, WI).
Hybridization protocol
Hybridization was performed by the Genomics and Bioinformatics Core Facility, University of Notre Dame, by using the Hybridization Kit, LS and Wash Buffer Kit (Roche NimbleGen, Madison, WI).
Scan protocol
Scanning was performed by the Genomics and Bioinformatics Core Facility, University of Notre Dame, using a NimbleGen MS 200 Microarray Scanner at 2 μM resolution.
Description
Wh_DD_4(2) hr constant darkness
Data processing
The raw data (.pair file) was subjected to RMA (Robust Multi-Array Analysis; Irizarry et al. Biostatistics 4(2):249), and background correction as implemented in the NimbleScan software package, version 2.4.27 (Roche NimbleGen, Inc.).
