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DNA Extraction Techniques for Genomic Analyses of Macroalgae

Wilson, Laura J.; Weber, Xénia A.; King, Tania M.; Fraser, Ceridwen

Description

Extracting high-quality DNA in large quantities from Phaeophyceae (brown), Rhodophyta (red), and Chlorophyta (green) macroalgae presents a substantial obstacle for modern molecular studies. Macroalgal tissues are rich in polysaccharides and polyphenols that are known to interfere with downstream molecular techniques. These compounds are released during DNA extraction procedures and often persist despite purification attempts. A wide range of DNA extraction and purification methods have...[Show more]

dc.contributor.authorWilson, Laura J.
dc.contributor.authorWeber, Xénia A.
dc.contributor.authorKing, Tania M.
dc.contributor.authorFraser, Ceridwen
dc.date.accessioned2018-06-29T03:57:34Z
dc.identifier.isbn978-94-017-7532-8
dc.identifier.urihttp://hdl.handle.net/1885/144648
dc.description.abstractExtracting high-quality DNA in large quantities from Phaeophyceae (brown), Rhodophyta (red), and Chlorophyta (green) macroalgae presents a substantial obstacle for modern molecular studies. Macroalgal tissues are rich in polysaccharides and polyphenols that are known to interfere with downstream molecular techniques. These compounds are released during DNA extraction procedures and often persist despite purification attempts. A wide range of DNA extraction and purification methods have been developed in attempts to overcome these challenges. Here, we review methods of macroalgal DNA extraction, including commercial kits. We discuss each method’s merits and limitations, and examine its potential use in both traditional and high-throughput molecular approaches. Finally, we present our own findings from a range of DNA extraction, purification, and sequencing trials based on the commercially available MoBio PowerPlant® Pro and PowerPlant® Clean kits, carried out on a number of Phaeophyceae species. We found that DNA yield and quality can be improved by lengthy soaking of tissue in extraction buffers, altering the homogenization procedures and modifying solutions used. The DNA extraction protocol presented here can be scaled up for high-throughput analyses and preliminary results suggest suitability for next-generation sequencing.
dc.description.sponsorshipLJW was funded by a scholarship from the North Australian Marine Research Association and an Australian Research Council Linkage grant led by Graham Edgar (University of Tasmania) (ARC LP100200122), XW was funded by a Capacity-Building scholarship from the Australian Biological Resources Study, TK was funded by the Department of Zoology at the University of Otago, and CIF was funded by an Australian Research Council DECRA grant (DE140101715).
dc.format.mimetypeapplication/pdf
dc.publisherSpringer Verlag
dc.relation.ispartofSeaweed Phylogeography
dc.rights© Springer Science+Business Media Dordrecht 2016
dc.subjectBrown algae
dc.subjectChlorophyta
dc.subjectGreen algae
dc.subjectHigh-throughput
dc.subjectInhibitors
dc.subjectPhaeophyceae
dc.subjectPolyphenols
dc.subjectPolysaccharides
dc.subjectRed algae
dc.subjectRhodophyta
dc.titleDNA Extraction Techniques for Genomic Analyses of Macroalgae
dc.typeBook chapter
dc.date.issued2016
local.publisher.urlhttps://link.springer.com
local.type.statusPublished Version
local.contributor.affiliationFraser, C. I., Fenner School of Environment and Society, The Australian National University
local.description.embargo2037-12-31
dc.relationhttp://purl.org/au-research/grants/arc/LP100200122
dc.relationhttp://purl.org/au-research/grants/arc/DE140101715
local.bibliographicCitation.startpage363
local.bibliographicCitation.lastpage386
local.identifier.doi10.1007/978-94-017-7534-2_15
dcterms.accessRightsOpen Access
CollectionsANU Research Publications

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