Table 1.

DNA extraction protocols of 3 optimized methods

Commercial DNA extraction (PureLink Genomic DNA kits)	DNAzol Direct extraction (Molecular Research Center, Catalog # DN131)	Microwave-based method
One female mosquito in a sterile 1.5-ml micro-centrifuge tube Add 20 μl of proteinase-K and 180 μl of lysis buffer, grind with micro-pestle for 1 min then vortex vigorously for 30 s Incubate at 55 °C, 2 h Centrifuge at 12,000 g for 3 min, transfer supernatant to a new sterile tube Add 20 µl RNase and incubate at room temperature for 2 min Add 200 μl of binding buffer and vortex FOR HOW LONG? Add 200 μl of absolute ethanol to lysate and mix gently for 5 s Transfer lysate to spin column in a collection tube and centrifuge at 12,000 g for 3 min Discard flow-through and then add 500 µl of washing buffer to the column tube Centrifuge at 12,000 g for 3 min Place the spin column into a new micro-centrifuge tube, then add 30 µl of elution buffer Centrifuge at 12,000 g for 5 min, then DNA keep at −20 °C	One female mosquito in a sterile 1.5-ml micro-centrifuge tube Add 150 µl of DNAzol Direct, grind with a sterile micro-pestle for 1 min then vortex vigorously for 30 s Incubate at 80 °C for 20 min Centrifuge at 12,000 g for 5 min, transfer supernatant to a new sterile tube Store supernatant at −20 °C	One female mosquito in a sterile 1.5-ml micro-centrifuge tube Add 200-μl sterile distilled water, grind with sterile micro-pestle for 1 min, then vortex vigorously for 30 s Heat in microwave at 800 W for 5 min Centrifuge at 12,000 g for 5 min, transfer supernatant to a new sterile tube Store supernatant at −20 °C