Utilize este identificador para referenciar este registo: http://hdl.handle.net/10400.1/4159
Título: Genomic DNA isolation from green and brown algae (Caulerpales and Fucales) for microsatellite library construction
Autor: Varela-Alvarez, E.
Andreakis, N.
Lago-Lestón, A.
Pearson, G. A.
Serrão, Ester
Procaccini, G.
Duarte, C. M.
Marbà, N.
Palavras-chave: Algae
Caulerpa
CTAB
DNA extraction
Microsatellite
Nuclei isolation
Population genetics
Sargassum
Seaweed
Data: Jun-2006
Editora: Phycological Society of America
Citação: Varela-Álvarez, E.; Andreakis, N.; Lago-Lestón, A.; Pearson, G.A.; Serrão, E.A.; Procaccini, G.; Duarte, C.M.; Marbá, N.Genomic DNA isolation from green and brown algae (Caulerpales and Fucales) for microsatellite library construction, Journal of Phycology, 42, 3, 741-745, 2006.
Resumo: A method for isolating high-quality DNA is presented for the green algae Caulerpa sp. (C. racemosa, C. prolifera, and C. taxifolia) and the brown alga Sargassum muticum. These are introduced, and invasive species in Europe, except for the native C. prolifera. Previous methods of extraction, using cetyl trimethyl ammonium bromide or various commercial kits, were used to isolate genomic DNA but either no DNA or DNA of very low quality was obtained. Genomic libraries were attempted with Caulerpa sp. on three occasions and either the restriction enzyme, the Taq polymerase, or the T4 ligase was inhibited, probably by the large amount of polysaccharides in these algae. The method presented here consists of the rapid isolation of stable nuclei, followed by DNA extraction. Yields of 6–10 μg genomic DNA from 1 g fresh blades were obtained. After genomic DNA was isolated from fresh material, the quality was checked by agarose gel. Quantification of DNA concentration was performed using UV spectrophotometric measurement of the A260/A280 ratio. The DNA was suitable for PCR, cloning, and hybridization. The DNA isolated using this method allowed successful construction of microsatellite libraries for Caulerpa species and S. muticum. The technique is inexpensive and appropriate for the isolation of multiple samples of DNA from a small amount of fresh material.
Peer review: yes
URI: http://hdl.handle.net/10400.1/4159
DOI: http://dx.doi.org/ 10.1111/j.1529-8817.2006.00218.x
ISSN: 0022-3646
Versão do Editor: http://onlinelibrary.wiley.com/doi/10.1111/j.1529-8817.2006.00218.x/abstract
Aparece nas colecções:CCM2-Artigos (em revistas ou actas indexadas)



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