At the crossroads of botanical collections and molecular genetics laboratory: testing methods to obtain amplifiable DNA from moss herbarium material
- Published
- Accepted
- Subject Areas
- Biodiversity, Molecular Biology, Plant Science
- Keywords
- CTAB DNA extraction, herbarium, Maritime Antarctic, mosses, Subantarctica, PCR, plant, Sanger sequencing, bryophytes
- Copyright
- © 2019 Saługa
- Licence
- This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, reproduction and adaptation in any medium and for any purpose provided that it is properly attributed. For attribution, the original author(s), title, publication source (PeerJ Preprints) and either DOI or URL of the article must be cited.
- Cite this article
- 2019. At the crossroads of botanical collections and molecular genetics laboratory: testing methods to obtain amplifiable DNA from moss herbarium material. PeerJ Preprints 7:e27904v1 https://doi.org/10.7287/peerj.preprints.27904v1
Abstract
Background. Museum collections, including herbarium specimens, are considered an invaluable source of DNA. They constitute a source of a precious commodity, particularly when it is difficult to obtain living material of rare species, or extant populations occurred only in hard to access geographical territories. It is apparent that herbaria should be directly linked with molecular genetics laboratories making them a quick, open-source for molecular projects. However, herbarium DNA is inherently characterised by high degradation and chemical modifications such as the presence of various secondary compounds. A wide range of DNA molecular techniques dedicated to the preserved plant material has been published so far. However, contrasting with a general interest in the application of molecular analyses in moss biology, no comprehensive assessment of DNA isolation and amplification methods from moss herbarium material, is available. Methods. To assess the feasibility of using DNA from moss herbarium specimens, we have tested and compared the silica column-based method and three variants of CTAB-based DNA extraction protocol. We used herbarium collections of twenty-five moss species collected between 1979 and 2013 and specifically focused on austral polar regions to assess the potential of herbarium as a source of biological material from geographical regions of difficult and restricted access. Results. Here, we present an optimized CTAB-based approach which effectively suppresses inhibitors in the herbarium DNA as was measured by amplification success. In this report, DNA purity and the length of the target genetic region are the fundamental agents which drive the successful PCR reaction. Conversely, the specimen age seems to be less relevant. Moreover, the size distribution of the DNA fragments extracted using Qiagen protocol is shown to be comparable to our original CTAB-based approach. Our modified CTAB-based method provides a high-purity genomic DNA allowing efficient downstream amplification. It is not outcompeted by the column-based method and appears as a method of choice in molecular studies of moss herbarium material.
Author Comment
This is a submission to PeerJ for review.