Comparison of Two Different DNA Isolation Methods (Quick & Dirty vs. Commercial Kit) in Small Insects
DOI:
https://doi.org/10.24925/turjaf.v12i10.1679-1683.6935Keywords:
COI, insect, classification, easy method, cheap method, species determinationAbstract
The COI gene region is the most commonly employed method for species identification in insects, as in all animals. The importance of DNA isolation methods is increasing, and they are of great importance for identifying insects. Unlike other large animals, DNA isolation from insects, especially from agricultural pests with very small structures, is possible with expensive commercial kits. In this study, an expensive commercial kit and a fast and economical method were compared in terms of time, cost, DNA quantity and quality. It was found that the success of commercial kits in terms of DNA quality and quantity is unquestionable. However, it was determined that the modified ‘quick and dirty’ method can be successfully used to extract DNA from insects and insect parts of all sizes, especially in studies that do not require long-term storage of DNA. Average isolation time for Q&D was 45 minutes, compared to 4 hours or more for commercial kits. Finally, cost is the most important factor in today’s global economy. The unit cost of DNA isolation using the Q&D method is approximately 88% lower.
References
Aravind, K., Ravikanth, G., Shaanker, R. U., Chandrashekara, K., Kumar, A. R. V., & Ganeshaiah, K. N. (2007). DNA barcoding: An exercise in futulity or utility? Current Science, 92(9), 1213-1216.
Ball, S.L., & Armstrong, K. F. (2008). Rapid, one-step DNA extraction for insect pest identification by using DNA barcodes. Journal of Economic Entomology, 101, 523-532. http://dx.doi.org/10.1093/jee/101.2.523
Castalanellia, M. A., Severtsonc, D. L., Brumleya, C. J., Szitoa, A., Foottitd, R. G., Grimma, M., Munyardb, K., & Grothb, D. M. (2010). A rapid non-destructive DNA extraction method for insects and other arthropods. Journal of Asia-Pacific Entomology, 3, 243-248.
Chen, H., Rangasamy, M., Tan, S. Y., Wang, H., & Siegfried, B. D. (2010). Evaluation of five methods for total DNA extractionfrom Western corn rootworm beetles. PLoSONE 5(8), 11963. doi:10.1371/journal.pone.0011963)
Favret, C. (2005). A new non-destructive DNA extraction and specimen clearing technique for aphids (Hemiptera). Proc. Enotomol. Soc. Wash. 107, 469–470.
Freitas, M.T., Gomes-Júnior, P.P., Batista, M.V., Leal-Balbino, T.C., Araujo, A.L., & Balbino, V.Q. (2014). Novel DNA extraction assay for molecular identification of Aedes spp. eggs. Genetics and Molecular Research, 13, 8776- 8782. http://dx.doi.org/10.4238/2014.
Gilbert, M. T. P., Moore, W., Melchior, L., & Worobey, M. (2007). DNA extraction from dry museum beetles without conferring external morphological damage. PloS One 2(3), e272. https://doi.org/10.1371/journal.pone.0000272
Hebert, P. D. N., & Gregory T. R. (2005). The promise of DNA barkoding for taxonomy. Systematic Biology, 54(5), 852-859. https://doi.org/10.1080/10635150500354886
Henry, J. M., Raina, A. K., & Ridgway, R. L. (1990). Isolation of High-Molecular-Weight DNA from insects. Analytical Biochemistry, 185, 147-150. http://dx.doi.org/10.1016/0003-2697(90)90270-J
Hunter, S.J., Goodall, T.I., Walsh, K. A., Owen, R., & Day, J. C. (2008). Nondestructive DNA extraction from blackflies (Diptera: Simuliidae): retaining voucher specimens for DNA barcoding projects. Molecular Ecology Resources, 8, 56–61. doi: 10.1111/j.1471-8286.2007.01879.x
Musapa, M., Kumwenda, T., Mkulama, M., Chishimba, S., Norris, D.E., Thuma, P.E., & Mharakurwa, S. (2013). A simple chelex protocol for DNA extraction from Anopheles spp. Journal of Visualized Experiments, 71, e3281.
Nancy, C. C., Mauricio, Q., Horacio, C. C., & Guadalupe, Z. P. (2010). A simple and rapid method for DNA isolation from xylophagous insects. International Journal of Molecular Sciences, 11, 5056-5064. http://dx.doi.org/10.3390/ijms11125056
Nelson, D. L., & Cox, M. M. (2005). Lehninger Principles of Biochemistry, Worth Publishers. ISBN: 1-57259-931-6
Pons, J. (2006). DNA-based identification of preys from non-destructive, total DNA extractions of predators using arthropod universal primers. Mol. Ecol. Notes, 6, 623–626.
Rowley, D. L., Coddington, J. A., Gates, M. W., Norrbom, A. L., Ochoa, R. A., Vandenberg, N. J., & Greenstone, M. H. (2007). Vouchering DNA-barcoded specimens: test of a nondestructive extraction protocol for terrestrial arthropods. Molecular Ecology Notes, 7, 915–924.
William, W. W., Mackey, K., & Chomczynski, P. (1997). Effect of pH and ionic strength on the spectrophotometric assessment of nucleic acid purity. BioTechniques, 22(3), 474-481. https://doi.org/10.2144/97223st01
Downloads
Published
How to Cite
Issue
Section
License
This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.
