Success of DNA extraction and PCR amplification from dry pinned sand bees (Andrena spp. Fabricius, 1775) using newly-designed primers

Abstract

The suitability of dry pinned museum specimens for DNA extraction of sand bees (Andrena spp. Fabricius, 1775) (Hymenoptera: Andrenidae) and the effectiveness of existing and new primers used in DNA analysis of specimens for future studies were evaluated. A total 256 specimens were analyzed, including 222 dry pinned bee specimens representing 37 subgenera and 101 species and 34 ethanol-preserved specimens belonging to 21 species. Several different protocols were tested for DNA extraction, and DNA was extracted from almost all of the specimens. The samples preserved in ethanol had the highest quality DNA. Of 31 primer sets tested for amplification of the DNA, 14 of them were newly designed or redesigned. The amplified sequence length ranged from 130 to 1571 bp. DNA from 32 specimens belonging to 25 species was successfully amplified at three to four loci. This study demonstrates the importance of storage conditions for specimens possibly destined for later DNA extraction, and for selecting suitable primers when dealing with older bee specimens. Some primers can be diagnostically informative provided appropriate gene regions are used.