Saginaw Valley State University, Biology, University Center, MI 48710
One of the biggest problems in entomology is species identification. However, every living thing comes with a built-in barcode written in the sequence of nucleotides in DNA. The Barcoding of Life Database (BOLD) project has as it's goal the acquisition of sequence data from every living species and has already acquired sequence data for over 36,000 species. The gene that has been chosen for the initial stage of the project for animals is the cytochrome oxidase subunit I (COI) gene of mitochondrial DNA. This gene has several characteristics that should make it useful for species identification and phylogenetic reconstruction at the species level. Two different primer combinations were chosen that allow for amplification of nearly 80% of the COI gene (Zhang and Hewitt, 1996, Insect Molecular Biology 6(2):143-150). DNA was extracted from 32 different species representing ten different orders. A 310 base pair fragment was successfully amplified from all 32 species using the primer combination UEA9/UEA10. A 1020 base pair fragment was successfully amplified from 22 species using primer combination UEA3/UEA8. Sequence data was obtained for seven species using the Beckman Coulter CEQ 8000 automated DNA sequencing system. The results indicate that it should be possible to obtain sequence data for most insect species quickly and cost-efficiently using fresh or frozen insect tissue. This method should serve as a powerful supplement to traditional taxonomic approaches.
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