Travis A Rainey, Emily E Tryc, Kirsten E Nicholson
{"title":"两栖动物基因采样中皮肤拭子、口腔拭子和脚趾夹的比较,以小型无尾目动物(acris blanchardi)为例进行研究","authors":"Travis A Rainey, Emily E Tryc, Kirsten E Nicholson","doi":"10.1093/biomethods/bpae030","DOIUrl":null,"url":null,"abstract":"\n Multiple methods for collecting genetic samples from amphibians exist, each with their own implications for study design, animal welfare, and costs. Toe clipping is one common method, but there is ongoing debate regarding its potential detriment. Less invasive methods should be implemented, if efficacious, as amphibians are a particularly vulnerable vertebrate group. Skin and buccal swabbing are less invasive methods for genetic sampling, but the potential for contamination and a lower yield of DNA may exist. To compare these methods, we gathered skin swabs, buccal swabs, and toe clips from the same individuals of a relatively small anuran species, Blanchard’s Cricket Frog (Acris blanchardi). We then compared DNA yield, DNA purity, amplification success rate, and genotypic data quality among sample types. We found toe clips and buccal swabs generated similar DNA yield and purity, with skin swabs yielding significantly less DNA of significantly lower purity than the other sample types. Amplification success rate was significantly higher using toe clips compared to the other sample types, though buccal swab samples amplified more readily than skin swabs. Genotypic data from toe clips and buccal swabs did not differ significantly in quality, but skin swab data quality was significantly lowest among sample types. Thus, skin swabbing could produce erroneous data in some situations, but buccal swabbing is likely an effective substitute to toe clipping, even for small species. Our results can help future researchers select which genetic sampling method might best suit their research needs.","PeriodicalId":36528,"journal":{"name":"Biology Methods and Protocols","volume":null,"pages":null},"PeriodicalIF":2.5000,"publicationDate":"2024-05-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"Comparing skin swabs, buccal swabs, and toe clips for amphibian genetic sampling, a case study with a small anuran (acris blanchardi)\",\"authors\":\"Travis A Rainey, Emily E Tryc, Kirsten E Nicholson\",\"doi\":\"10.1093/biomethods/bpae030\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"\\n Multiple methods for collecting genetic samples from amphibians exist, each with their own implications for study design, animal welfare, and costs. Toe clipping is one common method, but there is ongoing debate regarding its potential detriment. Less invasive methods should be implemented, if efficacious, as amphibians are a particularly vulnerable vertebrate group. Skin and buccal swabbing are less invasive methods for genetic sampling, but the potential for contamination and a lower yield of DNA may exist. To compare these methods, we gathered skin swabs, buccal swabs, and toe clips from the same individuals of a relatively small anuran species, Blanchard’s Cricket Frog (Acris blanchardi). We then compared DNA yield, DNA purity, amplification success rate, and genotypic data quality among sample types. We found toe clips and buccal swabs generated similar DNA yield and purity, with skin swabs yielding significantly less DNA of significantly lower purity than the other sample types. Amplification success rate was significantly higher using toe clips compared to the other sample types, though buccal swab samples amplified more readily than skin swabs. Genotypic data from toe clips and buccal swabs did not differ significantly in quality, but skin swab data quality was significantly lowest among sample types. Thus, skin swabbing could produce erroneous data in some situations, but buccal swabbing is likely an effective substitute to toe clipping, even for small species. Our results can help future researchers select which genetic sampling method might best suit their research needs.\",\"PeriodicalId\":36528,\"journal\":{\"name\":\"Biology Methods and Protocols\",\"volume\":null,\"pages\":null},\"PeriodicalIF\":2.5000,\"publicationDate\":\"2024-05-16\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Biology Methods and Protocols\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"https://doi.org/10.1093/biomethods/bpae030\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q3\",\"JCRName\":\"BIOCHEMICAL RESEARCH METHODS\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Biology Methods and Protocols","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.1093/biomethods/bpae030","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q3","JCRName":"BIOCHEMICAL RESEARCH METHODS","Score":null,"Total":0}
引用次数: 0
摘要
从两栖动物身上采集基因样本有多种方法,每种方法对研究设计、动物福利和成本都有各自的影响。剪趾是一种常见的方法,但关于这种方法的潜在危害一直存在争议。由于两栖动物是特别脆弱的脊椎动物群体,因此如果有效,应采用侵入性较小的方法。皮肤和颊拭子是侵入性较小的基因采样方法,但可能存在污染和 DNA 产量较低的可能性。为了比较这些方法,我们收集了皮肤拭子、口腔拭子和脚趾夹,这些拭子取自相对较小的无脊椎动物物种--布兰查德蟋蟀蛙(Acris blanchardi)的相同个体。然后,我们比较了不同样本类型的 DNA 产量、DNA 纯度、扩增成功率和基因型数据质量。我们发现脚趾夹和颊拭子产生的DNA产量和纯度相似,而皮肤拭子产生的DNA产量和纯度明显低于其他类型的样本。与其他样本类型相比,脚趾夹的扩增成功率明显更高,但颊拭子样本比皮肤拭子更容易扩增。趾夹和口腔拭子的基因型数据在质量上没有明显差异,但皮肤拭子的数据质量在所有样本类型中明显最低。因此,在某些情况下,皮肤拭子可能会产生错误的数据,但口腔拭子可能是剪趾的有效替代品,即使对于小型物种也是如此。我们的研究结果可以帮助未来的研究人员选择最适合其研究需要的基因采样方法。
Comparing skin swabs, buccal swabs, and toe clips for amphibian genetic sampling, a case study with a small anuran (acris blanchardi)
Multiple methods for collecting genetic samples from amphibians exist, each with their own implications for study design, animal welfare, and costs. Toe clipping is one common method, but there is ongoing debate regarding its potential detriment. Less invasive methods should be implemented, if efficacious, as amphibians are a particularly vulnerable vertebrate group. Skin and buccal swabbing are less invasive methods for genetic sampling, but the potential for contamination and a lower yield of DNA may exist. To compare these methods, we gathered skin swabs, buccal swabs, and toe clips from the same individuals of a relatively small anuran species, Blanchard’s Cricket Frog (Acris blanchardi). We then compared DNA yield, DNA purity, amplification success rate, and genotypic data quality among sample types. We found toe clips and buccal swabs generated similar DNA yield and purity, with skin swabs yielding significantly less DNA of significantly lower purity than the other sample types. Amplification success rate was significantly higher using toe clips compared to the other sample types, though buccal swab samples amplified more readily than skin swabs. Genotypic data from toe clips and buccal swabs did not differ significantly in quality, but skin swab data quality was significantly lowest among sample types. Thus, skin swabbing could produce erroneous data in some situations, but buccal swabbing is likely an effective substitute to toe clipping, even for small species. Our results can help future researchers select which genetic sampling method might best suit their research needs.