Matthew G Adair, Jean-Jacque Forgus, Devon C. Main, J. Taft, J. D. da Silva, K. Tolley
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引用次数: 0
Abstract
In biodiversity research, the retrieval of genetic material from organisms is a common and essential component for assessing genetic diversity. The welfare of the organism, however, needs to be balanced against the overall goal of the intended research. One sampling technique often applied to retrieve DNA material from small reptiles is the removal of a small portion of the distal end of the tail. While most squamate reptiles have tail autotomy, some species (e.g. many iguanid lizards and snakes) do not regenerate tail tissue. We therefore explored the efficacy of a minimally disruptive technique, buccal swabbing, as an alternative to tissue sampling via tail clipping, particularly for species without tail autotomy, using dwarf chameleons (Bradypodion spp.) as a case study. The two sampling techniques were compared to assess the efficacy of DNA retrieval. We also evaluated the financial implications of each technique. The results indicate that buccal swabs paired with a specialised DNA extraction kit offer a feasible (although expensive), once-off alternative to tissue sampling, but with no material left for biobanking. Deviations in swab type used and the DNA extraction process (i.e. using more affordable extraction procedures) resulted in poor DNA retrieval and unreadable sequences. This finding suggests that buccal swabbing can be a suitable alternative when finances are not constrained, an expensive extraction kit is available, and biobanking is not a concern. For researchers from low- to middle-income economies, this expensive alternative may hamper research progress by placing a financial obstacle in the way, and therefore the next best option is tissue sampling.
在生物多样性研究中,从生物体中获取遗传物质是评估遗传多样性的一个常见且重要的组成部分。然而,生物的福利需要与预期研究的总体目标相平衡。从小型爬行动物身上提取 DNA 材料经常采用的一种取样技术是切除尾巴远端的一小部分。虽然大多数有鳞类爬行动物都有尾部自体切除术,但有些物种(如许多巨蜥和蛇)的尾部组织不会再生。因此,我们以侏儒变色龙(Bradypodion spp.)我们对两种取样技术进行了比较,以评估 DNA 提取的效果。我们还评估了每种技术的财务影响。结果表明,颊拭子与专门的 DNA 提取试剂盒配对使用,是一种可行的一次性组织取样替代方法(尽管价格昂贵),但不会留下任何材料用于生物库。咽拭子类型和 DNA 提取过程的偏差(即使用更经济的提取程序)导致 DNA 提取效果不佳,序列无法读取。这一研究结果表明,当资金不紧张、有昂贵的提取试剂盒且不担心生物库问题时,颊拭子法不失为一种合适的替代方法。对于来自中低收入经济体的研究人员来说,这种昂贵的替代方法可能会因经济障碍而阻碍研究进展,因此下一个最佳选择是组织采样。
期刊介绍:
The South African Journal of Science is a multidisciplinary journal published bimonthly by the Academy of Science of South Africa. Our mandate is to publish original research with an interdisciplinary or regional focus, which will interest readers from more than one discipline, and to provide a forum for discussion of news and developments in research and higher education. Authors are requested to write their papers and reports in a manner and style that is intelligible to specialists and non-specialists alike. Research contributions, which are peer reviewed, are of three kinds: Review Articles, Research Articles and Research Letters.