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GENOMIC DNA ISOLATION FROM GREEN AND BROWN ALGAE ( CAULERPALES AND FUCALES ) FOR MICROSATELLITE LIBRARY CONSTRUCTION 1
Author(s) -
VarelaÁlvarez Elena,
Andreakis Nikos,
LagoLestón Asunción,
Pearson Gareth A.,
Serrão Ester A.,
Procaccini Gabriele,
Duarte Carlos M.,
Marbá Nuria
Publication year - 2006
Publication title -
journal of phycology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.85
H-Index - 127
eISSN - 1529-8817
pISSN - 0022-3646
DOI - 10.1111/j.1529-8817.2006.00218.x
Subject(s) - biology , genomic dna , dna extraction , genomic library , dna , microsatellite , polymerase chain reaction , microbiology and biotechnology , botany , biochemistry , gene , allele , base sequence
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 μ g 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 A 260 / A 280 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.