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Selective elimination of chloroplastidial dna for metagenomics of bacteria associated with the green alga caulerpa taxifolia (bryopsidophyceae)
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Orientador(es)
Resumo(s)
Molecular analyses of bacteria associated with
photosynthetic organisms are often confounded by
coamplification of the chloroplastidial 16S rDNA
with the targeted bacterial 16S rDNA. This major
problem has hampered progress in the characterization of bacterial communities associated to photo-synthetic organisms and has limited the full
realization of the potential offered by the last generation of metagenomics approaches. A simple and
inexpensive method is presented, based on ethanol
and bleach treatments prior to extraction, to efficiently discard a great part of chloroplastidial DNA
without affecting the characterization of bacterial
communities through pyrosequencing. Its effectiveness for the description of bacterial lineages associated to the green alga Caulerpa taxifolia (M. Vahl)
C. Agardh was much higher than that of the preexisting enrichment protocols proposed for plants.
Furthermore, this new technique requires a very
small amount of biological material compared to
the other current protocols, making it more realistic
for systematic use in ecological and phylogenetic
studies and opening promising prospects for
metagenomics of green algae, as shown by our data.
Descrição
Palavras-chave
16S Elimination Chloroplasts Metagenomics Green algae
Contexto Educativo
Citação
Aires, T.; Marbà, N.; Serrao, E.A.; Duarte, C.M.; Arnaud-Haond, S.Selective elimination of chloroplastidial dna for metagenomics of bacteria associated with the green alga caulerpa taxifolia (bryopsidophyceae), Journal of Phycology, 48, 2, 483-490, 2012
Editora
Phycological Society of America