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Refining DNA Barcoding Coupled High Resolution Melting for Discrimination of 12 Closely Related Croton Species
Chiang Mai Univ, Fac Sci, Dept Biol, Chiang Mai 50200, Thailand..
Chinese Acad Sci, Kunming Inst Zool, State Key Lab Genet Resources & Evolut, Kunming 650223, Peoples R China.;Chinese Acad Sci, Kunming Inst Zool, Yunnan Lab Mol Biol Domest Anim, Kunming 650223, Peoples R China.;Chiang Mai Univ, Sci & Technol Res Inst, Chiang Mai 50200, Thailand..
Chiang Mai Univ, Fac Sci, Dept Biol, Chiang Mai 50200, Thailand..
Chiang Mai Univ, Fac Sci, Dept Biol, Chiang Mai 50200, Thailand..
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2015 (English)In: PLoS ONE, ISSN 1932-6203, E-ISSN 1932-6203, Vol. 10, no 9, e0138888Article in journal (Refereed) Published
Abstract [en]

DNA barcoding coupled high resolution melting (Bar-HRM) is an emerging method for species discrimination based on DNA dissociation kinetics. The aim of this work was to evaluate the suitability of different primer sets, derived from selected DNA regions, for Bar-HRM analysis of species in Croton (Euphorbiaceae), one of the largest genera of plants with over 1,200 species. Seven primer pairs were evaluated (matK, rbcL1, rbcL2, rbcL3, rpoC, trnL and ITS1) from four plastid regions, matK, rbcL, rpoC, and trnL, and the nuclear ribosomal marker ITS1. The primer pair derived from the ITS1 region was the single most effective region for the identification of the tested species, whereas the rbcL1 primer pair gave the lowest resolution. It was observed that the ITS1 barcode was the most useful DNA barcoding region overall for species discrimination out of all of the regions and primers assessed. Our Bar-HRM results here also provide further support for the hypothesis that both sequence and base composition affect DNA duplex stability.

Place, publisher, year, edition, pages
2015. Vol. 10, no 9, e0138888
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Botany Evolutionary Biology
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URN: urn:nbn:se:uu:diva-265909DOI: 10.1371/journal.pone.0138888ISI: 000361800700111PubMedID: 26406615OAI: oai:DiVA.org:uu-265909DiVA: diva2:867071
Available from: 2015-11-04 Created: 2015-11-04 Last updated: 2017-12-01Bibliographically approved

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de Boer, Hugo

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