Keywords
molecular identification
museum collections
Hymenoptera
Coleoptera
Lepidoptera
How to Cite
Abstract
Insects constitute the most diverse group of animals on the planet, and their accurate identification is essential for research in conservation, agriculture, and taxonomy. In this context, DNA-based molecular techniques represent a highly valuable tool for their study; however, these tools are limited when working with specimens preserved in collections, as extraction protocols are often invasive and compromise the integrity of the insect body. The present study evaluated a minimally destructive DNA extraction protocol in two genera: Canthon sp. (Coleoptera) and Oxytrigona sp. (Hymenoptera). A total of 20 specimens were processed using a modification of the method described by Paydar et al. (2018). DNA quality was determined by spectrophotometry, measuring concentration and purity ratios A260/A280 and A260/A230. The results showed a markedly superior performance in Canthon sp., with concentrations of up to 563.2 ng/μl and purity ratios within the reference values, whereas Oxytrigona sp. yielded much lower concentrations (<3 ng/μl in most cases) and inadequate purity values. These differences can be attributed both to exoskeleton composition and to buffer formulation, as noted in previous studies. In conclusion, the adapted protocol proves effective for beetles, offering a non-destructive alternative for the molecular analysis of collection specimens, although adjustments are required for its application in hymenopterans.
References
Al-Dhafer, H. M., Balaji, R., Abdel-Dayem, M. S., Rasool, I., Mohamed, A., & Palanisamy, S. (2025). Simple DNA extraction for museum beetle specimens to unlock genetic data from historical collections. MethodsX, 14, 103236. https://doi.org/10.1016/j.mex.2025.103236
Amat-García, G., y Fernández, F. (2011). La diversidad de insectos (Arthropoda: Hexapoda) en Colombia I. Entognatha a Polyneoptera. Acta Biológica Colombiana, 16(2), 205–219. https:// www.redalyc.org/pdf/3190/319028008016.pdf
flDemeke, T., & Jenkins, G. R. (2010). Influence of DNA extraction methods, PCR inhibitors fiand quantification methods on real-time PCR assay of biotechnology-derived traits. Analytical and Bioanalytical Chemistry, 396(6), 1977–1990. https://doi.org/10.1007/ s00216-009-3150-9
Eggleton, P. (2020). The state of the world’s insects. Annual Review of Environment and Resources, 45, 61–82. https://doi.org/10.1146/ annurev-environ-012420-050035
Giordani, G., Whitmore, D., & Vanin, S. (2023). A new, non-invasive methodology for the molecular fiidentification of adult Sarcophagidae from collections. Insects, 14(7), 635. https://doi. org/10.3390/insects14070635
Korlević, P., McAlister, E., Mayho, M., Makunin, A., Flicek, P., & Lawniczak, M. K. N. (2021). A minimally morphologically destructive approach for DNA retrieval and whole-genome shotgun sequencing of pinned historic dipteran vector species. Genome Biology and Evolution, 13(10). https://doi.org/10.1093/gbe/evab226
Kaur, N., & Kaleka, A. S. (2022). Diversity, importance and decline of pollinating insects in present era. In Global decline of insects. IntechOpen. https://doi.org/10.5772/intechopen.100316
Liu, S., Sun, J., Yu, L., Zhang, C., Bi, J., Zhu, F., Qu, M., Jiang, C., & Yang, Q. (2012). Extraction and characterization of chitin from the beetle Holotrichia parallela Motschulsky. Molecules, 17(4), 4604–4611. https://doi.org/10.3390/ molecules17044604
Murthy, M. K., Khandayataray, P., Tara, M., Buragohain, P., Giri, A., & Gurusubramanian, G. (2022). Optimisation of DNA isolation and PCR techniques for beetle (Order: Coleoptera) specimens. International Journal of Tropical Insect Science, 42(3), 2761–2771. https://doi. org/10.1007/s42690-022-00736-3
Paydar, S., Ucar, E., Yari, P., Safavi, S. M., Kahrizi, D., Nateqi, M., Mirmoayedi, A., & Yari, K. (2018). fiA simplified and optimized protocol for total DNA extraction from insect species: Applicable for studying genetic diversity fiand PCR-based specimen identification fivia partial amplification of cytochrome oxidase I (COI) gene. Cellular and Molecular Biology, 64(12), 22. https://doi.org/10.14715/ cmb/2018.64.12.5
Patzold, F., Zilli, A., & Hundsdoerfer, A. K. (2020). Advantages of an easy-to-use DNA extraction method for minimal-destructive analysis of collection specimens. PLOS ONE, 15(7), e0235222. https://doi.org/10.1371/journal. pone.0235222
Promega. (2025). How do I determine the concentration, yield and purity of a DNA sample? https://www.promega.com/ resources /pubhub/enotes /how-do- i - determine-the-concentration-yield-and-purity-of-a-dna-sample/
Rizzo, D., Luchi, N., Da Lio, D., Bartolini, L., Nugnes, F., Cappellini, G., Bruscoli, T., Salemi, C., Griffo, R. V., Garonna, A. P., & Rossi, E. (2021). Development fiof a loop-mediated isothermal amplification fi(LAMP) assay for the identification of the invasive wood borer Aromia bungii (Coleoptera: Cerambycidae) from frass. 3 Biotech, 11(2). https://doi.org/10.1007/s13205- 020-02602-w
Waldschmidt, A. M., Fernandes Salomão, T. M., De Barros, E. G., & De Antônio Oliveira Campos, L. (1997). Extraction of genomic DNA from Melipona quadrifasciata (Hymenoptera: Apidae, Meliponinae). Brazilian Journal of Genetics, 20(3), 421–423. https://doi. org/10.1590/S0100-84551997000300011
Yang, R.-S., Ni, M.-Y., Gu, Y.-J., Xu, J.-S., Jin, Y., Zhang, J.- H., Wang, Y., & Qin, L. (2021). Newly emerging pest in China, Rhynchaenus maculosus (Coleoptera: Curculionidae): Morphology and fimolecular identification with DNA barcoding. Insects, 12(6), 568. https://doi.org/10.3390/ insects12060568
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