A procedure for combined genitalia dissection and DNA extraction in Lepidoptera
A procedure for combined genitalia dissection and DNA extraction in Lepidoptera
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DOI：
10.1163/187631204788912463
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发表时间：
2005-01-01
影响因子：
1.2
通讯作者：
Segerer, AH
中科院分区：
文献类型：
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作者：
Knölke, S;Erlacher, S;Segerer, AH
从鳞翅目昆虫中提取DNA是一个破坏性的过程，博物馆馆长通常不愿意提供博物馆标本进行分子研究。另一方面，解剖外生殖器和外生殖器是描述和鉴定物种的标准程序，即使是模式标本也被普遍接受。我们提出了一种方法，通过修改解剖协议的DNA序列信息的分析相结合的调查，在生殖器的形态特征。用酶消化软组织，然后提取DNA，代替了在氢氧化钾中浸渍纤维素。如线粒体COI基因所示，从黑腹果蝇中提取的DNA适合于测序，该基因适合于物种鉴定。经酶处理的大豆被证明足以保存形态特征。对采集标本的适当处理和酶消化的常规使用给出了建议。
Extraction of DNA from Lepidoptera is a destructive procedure and curators are often reluctant to provide museum specimens for molecular investigations. On the other hand, dissection of abdomens and genitalia is a standard procedure for description and identification of species and generally accepted even for type material. We present a method that combines the investigation of morphological traits in genitalia with the analysis of DNA sequence information by modifying the dissection protocol. Maceration of abdomens in potassium hydroxide is replaced by enzymatic digestion of soft tissue followed by DNA extraction. DNA extracted from abdomens is suitable for sequencing, as shown for the mitochondrial COI gene appropriate for species identification. Enzymatically treated abdomens proved to be sufficient for preservation of morphological traits. Recommendations are given for appropriate treatment of collected specimens and for routine use of enzymatic digestion.