A simple and cost-effective protocol for extraction of genomic DNA from ethanol preserved black flies(Simuliidae:Diptera)

Objective:To investigate the efficacy of extraction methodology for the proposed DNA from the ethanol preserved black flies(Simuliidae:Diptera).Methods:This study addressed a simple and effective protocol for extraction of DNA from black flies stored in the ethanol.The sizes of larval and adult black flies ranged from 1.5 to 2.5 mm and 3 to 7 mm,respectively.To demonstrate the efficacy of the proposed methodology,the DNA was extracted from the ethanol preserved sample of black flies using the commercial kids.The extracted DNA was further validated in the PCR amplification using internal transcribed spacer-1 rDNA and cytochrome oxidase subunit II rDNA target primers.Results:Interestingly,a minor modification in the proposed methodology yielded a good DNA concentration in comparison with the commercial kids.The extracted DNA sample using the proposed methodology was successfully validated in the PCR amplification using internal transcribed spacer-1 rDNA and cytochrome oxidase subunit II rDNA genes.Conclusions:The proposed DNA extraction procedure yielded good concentration of DNA from the ethanol preserved black flies.The added advantage is that the procedure is suitable for a range of insect species preserved in the ethanol obtained from the various field conditions.