Ringler Eva
Department of Integrative Biology and Physiology, University of California Los Angeles, 621 Charles E. Young Drive South, Los Angeles, CA 90095-1606, USA.
Department of Integrative Zoology, University of Vienna, Althanstrasse 14, A-1090 Vienna, Austria.
Amphib Reptil. 2018 Jan;39(2):245-251. doi: 10.1163/15685381-17000206. Epub 2018 Apr 24.
Skin swabbing, a minimally invasive DNA sampling method recently proposed for adult amphibians, was tested on the dendrobatid frog . I compared DNA yield from skin swabs and toe clips by evaluating obtained DNA concentrations and purity of extracts, as well as amplification success using eleven polymorphic microsatellite loci. I also tested whether storing skin swabs for two months at -20°C affected the properties of the extract or microsatellite analysis. Results show that skin swabs of adult suffered from high contamination and yielded significantly lower DNA quality and quantity, resulting in insufficient genotyping success, than DNA obtained from toe clips. The relatively dry skin in dendrobatid frogs may have impeded the collection of sufficient viable cells, and the presence of skin alkaloids and microbiota in the frog mucus may lead to high contamination load of skin swabs.
皮肤擦拭法是最近针对成年两栖动物提出的一种微创DNA采样方法，本研究在箭毒蛙身上对其进行了测试。我通过评估所获得的DNA浓度、提取物的纯度以及使用11个多态性微卫星位点的扩增成功率，比较了皮肤擦拭样本和趾尖样本的DNA产量。我还测试了将皮肤擦拭样本在-20°C下保存两个月是否会影响提取物的性质或微卫星分析。结果表明，成年箭毒蛙的皮肤擦拭样本污染严重，所产生的DNA质量和数量显著低于趾尖样本，导致基因分型成功率不足。箭毒蛙相对干燥的皮肤可能阻碍了足够数量活细胞的采集，并且青蛙黏液中存在的皮肤生物碱和微生物群可能导致皮肤擦拭样本的高污染负荷。
