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Research Project: Mitigating Alternate Bearing of Pecan - Bridge Project

Location: Fruit and Tree Nut Research

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

Author
item HAZIR, CANAN - Adnan Mederes University
item Bock, Clive

Submitted to: Turkish Journal of Entomology
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 2/27/2019
Publication Date: 3/26/2019
Citation: Hazir, C., Bock, C.H. 2019. Success rate of DNA extraction and PCR amplification from dry pinned sand bees (Andrena Fabricius, 1775) using newly - designed primers. Turkish Journal of Entomology. 43:79-96. https://doi.org/10.16970/entoted.460948.
DOI: https://doi.org/10.16970/entoted.460948

Interpretive Summary: The suitability of dry pinned museum specimens for DNA extraction from sand bees (Hymenoptera) and the effectiveness of existing and new primers used in DNA analysis of specimens for future studies of their genetics was evaluated. A total 254 specimens were analyzed. Different protocols were tested for DNA extraction. Some samples preserved in ethanol were included and had the highest quality DNA. Of 31 primer sets tested for DNA amplification, 14 were newly designed or redesigned. DNA from only 32 specimens was successfully amplified at three to four loci. The study demonstrates the importance of storage conditions for specimens where DNA extraction is required, and for selecting suitable primers when dealing with older bee specimens. Some primers may be diagnostically informative provided appropriate gene regions are used.

Technical Abstract: The suitability of dry pinned museum specimens for DNA extraction of sand bees (Andrena 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 254 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.