A partial COI was amplified at locations 1751–2191 on the basis of
the published Drosophila yakuba mtDNA sequence (Clary and Wolstenholme, 1985).
PCR was performed using a diluted gDNA sample
(50–100 times with deionized water) along with primers of C1-J-1751
(5′-GGATCACCTGATATAGCATTCCC-3′) and C1-N-2191 (5′CCAGGTAAAATTAAAATATAAACTTC-3′) for 35 cycles of 1 min at 94°C, 45 s at 50 °Cand 1 min 30 s at 72 °C. The resulting PCR product was treated with PCR
quick-spin™(Intron Biotechnology, Seongnam, Korea) to remove small
nucleic acids and to replace PCR buffer with deionized water.