A rapid method of non-destructive DNA extraction from individual springtails (Collembola)

被引:13
|
作者
Aoyama, Hiroaki [1 ]
Saitoh, Seikoh [1 ]
Fujii, Saori [2 ]
Nagahama, Hideki [1 ]
Shinzato, Naoya [1 ]
Kaneko, Nobuhiro [2 ]
Nakamori, Taizo [2 ]
机构
[1] Univ Ryukyus, Trop Biosphere Res Ctr, Nishihara, Okinawa 9030213, Japan
[2] Yokohama Natl Univ, Dept Environm & Nat Sci, Grad Sch Environm & Informat Sci, Yokohama, Kanagawa 2408501, Japan
关键词
Collembola; DNA barcoding; Non-destructive extraction; Single-individual PCR; Voucher; SPECIMENS; AMPLIFICATION; BARCODE;
D O I
10.1007/s13355-015-0340-0
中图分类号
Q96 [昆虫学];
学科分类号
摘要
In this study, we describe an easy and rapid method for non-destructive DNA extraction from a single Collembola individual, without dissection, lysis of the specimen, or purification of extracted DNA. We demonstrate that, after a single specimen has been heat-treated in Tris-EDTA (TE) buffer using a standard thermocycler, the solution can be used for PCR amplification of the mitochondrial cytochrome c oxidase subunit 1 (COI) gene region, typically used for DNA barcoding. With this method, the morphological features of Collembola commonly used for species identification are well preserved. This DNA extraction method is preferable for DNA barcoding where the sequencing and preservation of a large number of small and fragile specimens are required.
引用
收藏
页码:419 / 425
页数:7
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