Pooled template whole genome sequencing (PoolSeq) of highly resistant Anopheles funestus mosquitoes that survived exposure to 1×, 5×, and 10× the diagnostic concentration of pyrethroids, as well as genetically crossed strains, in Mibellon, Cameroon.
Pooled template whole genome sequencing (PoolSeq) was conducted using Anopheles funestus (sensu stricto) mosquitoes that survived exposure to 1×, 5×, and 10× doses of pyrethroids, as well as highly resistant genetic crosses generated in the laboratory. The mosquitoes used in this study were field-caught females from Mibellon, Cameroon.  Indoor-resting female mosquitoes were collected and reared, and F1 progeny were exposed to pyrethroids to discriminate between live and dead phenotypes at each concentration. Genetic crosses were also established and phenotyped. Total genomic DNA was then extracted from individual mosquitoes using the QIAGEN DNeasy Blood and Tissue Kit (Qiagen, Hilden, Germany) following the manufacturer's instructions. Extracted genomic DNA was quantified per individual and pooled in equal amounts to form sequencing pools (n = 40 in all cases). For each pool, whole-genome DNA sequencing (DNA-seq) libraries were prepared using the TruSeq Nano (Low-Throughput) DNA Library Preparation Kit (Illumina). Libraries were pooled in equimolar amounts, and sequencing was performed on an Illumina NovaSeq 6000 platform using 2×150 bp paired-end sequencing with v4 chemistry.