松墨天牛成虫标本保存及其DNA提取质量比较
Comparison of the quality of genomic DNA extracted from adult specimens of Monochamus alternatus preserved by different methods
曲良建1** 王丽娟2 王青华1 王玉珠1 张永安1***
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DOI：
作者单位：1. 中国林业科学研究院森林生态环境与保护研究所 北京 100091；2. 中国林业科学研究院林业研究所 北京 100091
中文关键词：松墨天牛，天牛成虫，DNA提取，保存方式
英文关键词： Monochamus alternates, longicorn adult, DNA extraction, preservation methods
中文摘要： 【目的】 探讨适合DNA 提取的天牛成虫标本保存方法。【方法】 采用SDS-蛋白酶K消化法对液氮中冷冻保存、无水乙醇－20℃冷冻保存、无水乙醇室温保存和干标本室温保存且保存时间在2年以上的松墨天牛Monochamus alternates Hope成虫标本基因组DNA进行提取，并对不同保存方式提取的DNA样本进行了质量比较和分析。【结果】 在上述常见的松墨天牛成虫标本4种保存方式中，以液氮中冷冻保存效果最佳，其次为无水乙醇－20℃冷冻保存，插针干标本室温保藏效果最差。利用昆虫线粒体基因COⅠ和COⅡ的通用引物从上述DNA中均能够成功扩增出目的片段，测序结果证实扩增片段符合预期。【结论】 液氮和无水乙醇－20℃冷冻保存适合松墨天牛成虫标本长期保存，且不影响后续的PCR扩增和测序。
英文摘要： [Objectives] To find which method of preserving Monochamus alternatus specimens produced the best quality genomic DNA. [Methods] Genomic DNA from Monochamus alternatus adults that had been preserved using different methods (liquid nitrogen, 100% ethanol at -20℃, 100% ethanol at room temperature and museum specimens) for more than 2 years, was extracted using the SDS-proteinase K method and the quality of the extracted genomic DNA compared. [Results] The best quality genomic DNA was obtained from specimens that had been stored in liquid nitrogen, and the next best from those that had been preserved in 100% ethanol at -20℃. [Conclusion] The best quality DNA was extracted from specimens that had been frozen in liquid nitrogen or preserved in 100% ethanol at -20℃. The DNA samples obtained from such specimens are suitable for PCR amplification and sequencing.