A note on DNA identification of mosquito specimens for long-term preservation

Abstract

The cytochrome oxidase subunit I (COI) region of mitochondrial DNA (mtDNA) and internal transcriber spacer 2 (ITS2) ribonuclear DNA were used to amplify specific regions from total genomic DNA of preserved Anopheles minimus Theobald collected from Ratchaburi Province, Thailand. The universal primers were used to compare the techniques for long-term preservation. The experiments were conducted using the offspring of each individual mosquito. Mosquitoes were reared for F1 generation which then preserved as frozen and dried specimens. Frozen mosquitoes yielded amplified PCR product for both COI and ITS2 regions, whereas dried preserved mosquitoes yielded PCR product for only ITS2 region. The comparison of DNA recovery by PCR amplification of preserved techniques for dried mosquitoes, stored for up to 22 years, showed that pin steel specimens were more efficient than paper-point specimens in the way that paper-point technique needed the nail Polisher for attachment. This polisher might cause damage of DNA from different specimen preservations should be considerable in order to select the method of preservations for specific purposes and advance uses of these specimens in the future.