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| Status |
Public on May 12, 2015 |
| Title |
No.6 resistant housefly strain |
| Sample type |
SRA |
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| Source name |
791 resistant strain
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| Organism |
Musca domestica |
| Characteristics |
strain: 791spin
insecticide resistance: Spinosad
age: 5-7 days
gender: pooled male and female
developmental stage: adult
tissue: whole fly
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| Growth protocol |
The spinosad selected 791spin strain was created by selection with spinosad of the multi-resistant 791a strain strain that was collected in Denmark in 1997. The 791spin females are 21-fold spinosad-resistant to spinosad at the LC50, whereas 791spin male houseflies were 6-fold resistant. The flies were collected on private land with consent of the owner. The field collection did not involve endangered or protected species.Housefly breeding followed standard laboratory conditions. Egg laying was performed on crumpled filter paper soaked in whole milk. Breeding jars (5 L plastic buckets) containing 4 L of medium were seeded with 200 mg of eggs, corresponding to 2700 eggs. The breeding medium consisted of wheat bran 400 g, lucerne meal 200 g, baker’s yeast 10 g, malt extract 15 mL, whole milk 500 mL and water 500 mL. For adult feeding, cube sugar and water were given continuously. Feeding started after emergence with whole-milk powder mixed with icing sugar (1:1 w/w).Houseflies for transcriptome analysis were five to seven days old, adult male and female flies, which were fed sugar as the only food source.
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| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA from whole bodies of pooled flies (approx. 1.2 g equivalent to 60 flies) was extracted using the RNeasy Maxi Kit (Qiagen). Flies were thoroughly ground with liquid nitrogen, a mortar and pestle and otherwise following the manufacturer’s protocol. Isolated RNA was DNase-treated and concentrated using the RNeasy MinElute Kit (Qiagen). Gel electrophoresis and spectrophotometry (Nanodrop; NanoDrop Technologies, Wilmington, USA) was performed to assess the integrity and the concentration of each RNA sample, which was dissolved in RNase-free water and stored at -20ºC until use.
A normalized cDNA library was prepared from 12.2 µg mRNA prepared from adult male and female houseflies. The normalized cDNA library was size fractioned to approx. 500-1,200 bp. High throughput sequencing on GS FLX Titanium of the Musca cDNA library was done according to the standard protocols using a Genome Sequencer FLX Titanium Instrument (Roche Diagnostics). Preparation of cDNA, normalization and sequencing was performed by Eurofins MWG GmbH (Ebersberg, Germany).
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| Library strategy |
RNA-Seq |
| Library source |
transcriptomic |
| Library selection |
cDNA |
| Instrument model |
454 GS FLX Titanium |
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| Data processing |
After removal of adaptor sequences, data were aligned and de novo assembled using version 2.6 of the newbler assembler (454 Life Sciences/Roche, Branford. Clustering and assembly of all reads in contigs after the sequencing were done using MIRA 4.0 and contigs were initially analyzed by BLAST analysis.
Genome_build: de novo assembly
Supplementary_files_format_and_content: Total and normalized counts
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| Submission date |
Feb 12, 2015 |
| Last update date |
May 15, 2019 |
| Contact name |
Khalid Mahmood |
| E-mail(s) |
khalidwattoecol@gmail.com
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| Phone |
4542783010
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| Organization name |
Aarhus University
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| Department |
Agro-ecology
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| Lab |
Molecular Ecology
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| Street address |
Forsøgsvej 1
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| City |
Slagelse |
| ZIP/Postal code |
DK-4200 |
| Country |
Denmark |
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| Platform ID |
GPL19777 |
| Series (1) |
| GSE65891 |
Transcriptome analysis of insecticide resistant housefly strain: insights about SNPs and regulatory elements in differentially expressed cytochrome P450 genes |
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| Relations |
| BioSample |
SAMN03342161 |
| SRA |
SRX876109 |
| Supplementary data files not provided |
SRA Run Selector |
| Raw data are available in SRA |
| Processed data are available on Series record |
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