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タイトル: Methods for retaining well-preserved DNA with dried specimens of insects
著者: NAKAHAMA, Naoyuki
ISAGI, Yuji  KAKEN_id  orcid https://orcid.org/0000-0002-9777-076X (unconfirmed)
ITO, Motomi
著者名の別形: 中濵, 直之
井鷺, 裕司
伊藤, 元己
キーワード: Orthoptera
Gryllidae
Acheta domestica
DNA fragmentation
dried specimens of insects
genetic analysis
propylene glycol
発行日: 2019
出版者: Institute of Entomology, Biology Centre, Czech Academy of Sciences
誌名: European Journal of Entomology
巻: 116
開始ページ: 486
終了ページ: 491
抄録: Dried specimens of insects are increasingly seen as genetic resources. However, genetic analysis of dried specimens of insects is hampered by the deterioration of the DNA. In this study, we developed methods for preparing dried specimens of insects with well-preserved DNA, mainly for PCR-based genetic analysis. First, we compared the effects of either exposure to ethyl acetate vapour for from 10 min to 6 h or by freezing on the fragmentation of DNA in order to determine optimal length of time needed for killing insects using the above methods. Second, we compared the fragmentation of DNA after preservation by drying or immersion of legs in 99.5% ethanol or 99% propylene glycol in 0.2-ml tubes. We assessed degrees of fragmentation of DNA by determining polymerase chain reaction (PCR) success rates with primers for 313-, 710- and 1555-bp fragments using DNA that was collected immediately, and at one, six and 12 months after preparing the specimens. Differing times taken to kill insects did not affect the fragmentation of DNA. In dried specimens, DNA was seriously fragmented after one month, whereas that in legs prepared by immersion in 99.5% ethanol or 99% propylene glycol contained long fragments of DNA (1555 bp~) after 12 months. Propylene glycol was more suitable for preservation than ethanol, because the latter evaporates. Thus, to preserve insect DNA we suggest inserting the pin on which an insect is impaled into the hinged lid of a 0.2-ml tube containing 99% propylene glycol so that when the lid is closed the legs of the insect are preserved in the solution.
記述: 遺伝情報を長期保存できる昆虫標本の作製方法を新たに開発 --「遺伝資源」としての昆虫標本を次世代に--. 京都大学プレスリリース. 2019-12-26.
著作権等: © Institute of Entomology, Biology Centre, Czech Academy of Sciences, České Budějovice. This is an open access article distributed under the terms of the Attribution 4.0 International (CC BY 4.0), which permits use, distribution, and reproduction in any medium, provided the original publication is properly cited. No use, distribution or reproduction is permitted which does not comply with these terms.
URI: http://hdl.handle.net/2433/245245
DOI(出版社版): 10.14411/eje.2019.050
関連リンク: https://www.kyoto-u.ac.jp/ja/research-news/2019-12-26
出現コレクション:学術雑誌掲載論文等

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