A rapid method of non-destructive DNA extraction from individual springtails (Collembola)
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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.
KeywordsCollembola DNA barcoding Non-destructive extraction Single-individual PCR Voucher
We thank Kamigamo Experimental Station, Field Science Education and Research Center (FSERC), Kyoto University, Japan, for allowing access to the study site. We are grateful to Dr. K. Ichisawa at the Tottori Prefectural Museum, Tottori, for depositing our specimens in the museum. We thank two anonymous reviewers for their careful reading of the manuscript and constructive comments. This study was partly supported by MEXT/JSPS KAKENHI grant no. 25281029.
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