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Simple and rapid RNA extraction from freeze-dried tissue of brown algae and seagrasses

2006-02Journal article Restricted access
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dc.contributor.authorPearson, G. A.
dc.contributor.authorLago-Lestón, Asunción
dc.contributor.authorValente, M.
dc.contributor.authorSerrão, Ester
dc.date.accessioned2014-06-03T13:54:27Z
dc.date.available2014-06-03T13:54:27Z
dc.date.issued2006-02
dc.date.updated2014-05-21T13:41:52Z
dc.description.abstractAn 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.por
dc.identifier.citationPearson, 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.por
dc.identifier.doihttp://dx.doi.org/ 10.1080/09670260500505011
dc.identifier.issn0967-0262
dc.identifier.otherAUT: ESE00527;
dc.identifier.urihttp://hdl.handle.net/10400.1/4169
dc.language.isoengpor
dc.peerreviewedyespor
dc.publisherTaylor & Francispor
dc.relation.publisherversionhttp://www.tandfonline.com/doi/abs/10.1080/09670260500505011#.U43Sn3JdWxopor
dc.subjectBrown algaepor
dc.subjectFucuspor
dc.subjectLyophilizationpor
dc.subjectPlastid gene expressionpor
dc.titleSimple and rapid RNA extraction from freeze-dried tissue of brown algae and seagrassespor
dc.typejournal article
dspace.entity.typePublication
oaire.citation.endPage104por
oaire.citation.issue1por
oaire.citation.startPage97por
oaire.citation.titleEuropean Journal of Phycologypor
oaire.citation.volume41por
person.familyNamePearson
person.familyNameLago-Lestón
person.familyNameSerrao
person.givenNameGareth Anthony
person.givenNameAsunción
person.givenNameEster A.
person.identifier113536
person.identifierC-6686-2012
person.identifier.ciencia-id3315-9919-1A52
person.identifier.ciencia-id5B13-B26E-B1EC
person.identifier.orcid0000-0002-0768-464X
person.identifier.orcid0000-0003-4361-7732
person.identifier.orcid0000-0003-1316-658X
person.identifier.ridJ-3911-2013
person.identifier.scopus-author-id55916875600
person.identifier.scopus-author-id57193972525
person.identifier.scopus-author-id7004093604
rcaap.rightsrestrictedAccesspor
rcaap.typearticlepor
relation.isAuthorOfPublication0a10c448-c7f5-4c3b-9488-3917c707e35e
relation.isAuthorOfPublicationcf3375ea-836f-4de2-9cd6-fde518b26838
relation.isAuthorOfPublication45ccfe90-155c-4d6f-9e86-8f0fd064005f
relation.isAuthorOfPublication.latestForDiscovery0a10c448-c7f5-4c3b-9488-3917c707e35e

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