Reviving Galápagos snails: ancient DNA extraction and amplification from shells of probably extinct endemic land snails
Reviving Galápagos snails: ancient DNA extraction and amplification from shells of probably extinct endemic land snails
复制标题
复活加拉帕戈斯蜗牛：从可能灭绝的特有蜗牛壳中提取和扩增古代 DNA
DOI：
10.1093/mollus/eyw011
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发表时间：
2016
影响因子：
1.2
通讯作者：
B. Kemp
中科院分区：
文献类型：
--
作者：
Fernando A. Villanea;C. Parent;B. Kemp
蜗牛壳代表了关于建造它们的生物体的丰富信息来源，这对于在当地或全球灭绝的物种特别重要和相关。从蜗牛壳中获取遗传信息可能是有价值的，但以前从空壳中提取DNA的方法成功率极低，特别是从长期暴露于环境条件下风化的壳中提取。在这里，我们提出了两个简单的协议提取和扩增的DNA从空的土地蜗牛壳标本加拉帕戈斯特有的蜗牛，包括可能灭绝的物种。我们处理了来自加拉帕戈斯群岛的35个Naesiotus属（Bulimulidae）的贝壳，其中一些来自过去50年中没有观察到的物种。对18例正常人的线粒体DNA（mtDNA）进行了扩增和序列测定。我们的研究结果表明，实施一个古老的DNA提取协议和仔细的引物设计，以目标短的DNA片段，可以导致成功恢复mtDNA数据从这样的标本。
Snail shells represent an abundant source of information about the organisms that build them, which is particularly vital and relevant for species that are locally or globally extinct. Access to genetic information from snail shells can be valuable, yet previous protocols for extraction of DNA from empty shells have met with extremely low success rates, particularly from shells weathered from long-term exposure to environmental conditions. Here we present two simple protocols for the extraction and amplification of DNA from empty land snail shells from specimens of Galápagos endemic snails, including presumably extinct species. We processed 35 shells of the genusNaesiotus(Bulimulidae) from the Galápagos islands, some from species that have not been observed alive in the past 50 years. We amplified and sequenced short fragments (≤244 bp) of mitochondrial DNA (mtDNA) from 18 specimens. Our results indicate that the implementation of an ancient DNA extraction protocol and careful primer design to target short DNA fragments can result in successful recovery of mtDNA data from such specimens.