Genomic DNA was extracted from eight individual female mosquitoes using the MagExtractor Genome kit (Toyobo Co. Ltd., Osaka, Japan) (52 (link)). Index library construction and targeted capture of pooled libraries were conducted as previously described (52 (link)). Illumina library construction and hybridization capture was conducted with the biotinylated oligo probe designed from the A. albopictus Vgsc gene, whose exons show >92.5% homologies to the exons in A. aegypti (52 (link)). The quantified library was sequenced using the Illumina MiniSeq with the Mid Output Kit (Illumina Inc., San Diego, CA) and 151 cycles for both ends. Read pairs of 114 to 349 kbp were sequenced for each sample. Raw fastq read data were deposited to the National Center for Biotechnology Information (NCBI) Sequence Read Archive (BioProject ID: PRJNA795523). NGS-based reads were mapped to the reference Vgsc genome sequence (AALF000723-RA) and annotated for the synonymous and nonsynonymous nucleotide polymorphisms by using the automated MoNas pipeline (https://github.com/ItokawaK/MoNaS ) as previously described (52 (link)). Other detailed information about NGS analysis with target capture probes is described in our previous paper (52 (link)).
Protocol detail
Targeted Sequencing of Mosquito Insecticide Resistance Gene
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Corresponding Organization : National Institute of Infectious Diseases
Other organizations : National Agriculture and Food Research Organization, National Institute of Animal Health, Nagasaki University, National Institute Of Hygiene And Epidemiology, Royal University of Agriculture, Cambodia, The University of Tokyo, National Environment Agency, Nanyang Technological University, Ministry of Health and Welfare, Noguchi Memorial Institute for Medical Research, University of Ghana, Airlangga University
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Variable analysis
independent variables
- Genomic DNA extraction method (MagExtractor Genome kit)
- Mosquito samples (eight individual female mosquitoes)
- Nucleotide polymorphisms in the Vgsc gene
- Biotinylated oligo probe designed from the A. albopictus Vgsc gene, whose exons show >92.5% homologies to the exons in A. aegypti
- Positive control: Not explicitly mentioned
- Negative control: Not explicitly mentioned
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