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Data from: High-throughput genotyping of Anopheles mosquitoes using intact legs by Agena Biosciences iPLEX

Citation

Fabrigar, Danica Joy; Hubbart, Christina; Miles, Alistair; Rockett, Kirk (2015), Data from: High-throughput genotyping of Anopheles mosquitoes using intact legs by Agena Biosciences iPLEX, Dryad, Dataset, https://doi.org/10.5061/dryad.nr716

Abstract

Recent developments in genotyping technologies coupled with the growing desire to characterise genome variation in Anopheles populations opens the opportunity to develop more effective genotyping strategies for high-throughput screening. A major bottleneck of this goal is nucleic acid extraction. Here, we examined the feasibility of using intact portions of a mosquito's leg as sources of template DNA for whole genome amplification (WGA) by Primer-Extension Pre-amplification. We used the Agena Biosciences MassARRAY platform (formerly Sequenom) to genotype 78 SNPs for 265 WGA leg samples. We performed nucleic acid extraction on 36 mosquito carcasses and compared the genotype call concordance with their corresponding legs, and observed full concordance. Using three legs instead of one improved genotyping success rates (96% versus 89%, respectively), although this difference was not significant. We provide a proof of concept that WGA reactions can be performed directly on mosquito legs, thereby eliminating the need to extract nucleic acid. This approach is straightforward, sensitive and allows both species determination and genotyping of Anopheles mosquitoes to be performed in a high-throughput manner. Our protocol also leaves the mosquito body intact facilitating other experimental analysis to be undertaken on the same sample. Based on our findings, this method would also be suitable for use with other insect species.

Usage Notes

Location

Uganda