Using eight universal primers and new designed 315 species-specific primers, we tried to retrieve COI sequences from 45 dried specimens of 36 butterfly species collected from 1959 to 1980. The eight universal primers were entirely failed in PCR amplification and sequencing of all specimens. In the other hand, the 315 primers, targeting fragments of 71–417 bp, generated various lengths of COI sequences ranged from 444 bp to 658 bp from all specimens. Among 284 primer pairs, 26 primer pairs designed for Limenitis camilla, Argynnis niobe, and Brenthis daphne were success to produce COI sequences of congeneric speices, Limenitis doerriesi, Argynnis nerippe, and Brenthis ino. It suggests that the species-specific primers can be applied for analyzing COI sequences of closely related species. Our study reveals that newly designed species-specific primers will be effective to retrieval of COI sequences of old butterfly specimens.

• Taeman Han(Applied Entomology Division, Department of Agricultural Biology, National Academy of Agricultural Science, Suwon, Korea)
• Wonhoon Lee(Animal and Plant Quarantine Agency, Busan, Korea)
• Youngbo Lee(Applied Entomology Division, Department of Agricultural Biology, National Academy of Agricultural Science, Suwon, Korea)
• Namjung Kim(Applied Entomology Division, Department of Agricultural Biology, National Academy of Agricultural Science, Suwon, Korea)
• Shin-ichi Akimoto(Lap of Systematic Entomology, graduate School of Agriculture, Hokkaido University, Japan)
• Haechul Park(Applied Entomology Division, Department of Agricultural Biology, National Academy of Agricultural Science, Suwon, Korea)