Utilize este identificador para referenciar este registo: http://hdl.handle.net/10400.1/4169
Título: Simple and rapid RNA extraction from freeze-dried tissue of brown algae and seagrasses
Autor: Pearson, G. A.
Lago-Lestón, A.
Valente, M.
Serrão, Ester
Palavras-chave: Brown algae
Plastid gene expression
Data: Fev-2006
Editora: Taylor & Francis
Citação: Pearson, G.A.; Lago-Leston, A.; Valente, M.; Serrão, E.A.Simple and rapid RNA extraction from freeze-dried tissue of brown algae and seagrasses, European Journal of Phycology, 41, 1, 97-104, 2006.
Resumo: An inexpensive and rapid RNA extraction protocol for brown algae and seagrasses is presented, based on homogenization in a simple CTAB buffer and selective precipitation of RNA with lithium chloride. The protocol avoids the use of toxic chaotropic agents and phenol; high concentrations of dithiothreitol are used to inhibit RNase activity and to prevent oxidative cross-linking of nucleic acids by phenolics. A relatively high throughput of about 100 samples in 24 h can be achieved for, for example, Northern analysis. Yields of 50–200 µg g−1 fresh weight are comparable with those obtained for higher plants using commercially available kits. Tests of the extraction procedure demonstrate that high quality, intact RNA can be obtained from a variety of lyophilized brown algal tissues, even after prolonged storage at room temperature. Lyophilization is therefore suggested as an alternative to freezing tissue at −70°C to −80°C. The RNA obtained was used directly in several downstream applications to detect Fucus plastid-encoded transcripts by RNA-labelling, RT-PCR and Northern analysis.
Peer review: yes
URI: http://hdl.handle.net/10400.1/4169
DOI: http://dx.doi.org/ 10.1080/09670260500505011
ISSN: 0967-0262
Versão do Editor: http://www.tandfonline.com/doi/abs/10.1080/09670260500505011#.U43Sn3JdWxo
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