A small-scale RNA isolation protocol useful for high-throughput extractions from recalcitrant plants

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dc.contributor.authorSmart, Mariette
dc.contributor.authorRoden, Laura Catherine
dc.date.accessioned2018-05-11T14:23:59Z
dc.date.available2018-05-11T14:23:59Z
dc.date.issued2010
dc.date.updated2016-01-13T11:06:14Z
dc.description.abstractMany plants indigenous to South Africa are rich in secondary and oxidizing compounds such as pigments, complex polysaccharides and polyphenols. This makes isolation of high quality RNA for analysis of gene expression difficult. Here we describe a cost-effective isolation protocol suitable for RNA extraction from recalcitrant plant species. This method uses small amounts of tissue, so is useful when material is limited, and is easy to process large numbers of samples at once. We have used the method successfully with mature leaves of Protea hybrid ‘Sylvia’, and species P. repens, Leucospermum hybrid ‘Succession’, resurrection plants Xerophyta humilis and Craterostigma pumilum, and mature needles of Pine (Pinus radiata). RNA was analyzed spectrophotometrically and was found to be of high purity with low levels of contaminating compounds. Electrophoretic analyses on denaturing formaldehyde agarose gels and an Agilent 2100 Bioanalyzer confirmed the presence of RNA of high integrity. This is the first description of plant RNA integrity number (RIN) values for these plants using the algorithm designed for analyses of plant RNA containing multiple ribosomal bands. The RNA could successfully be used for reverse transcription and gene amplification.
dc.identifierhttp://dx.doi.org/10.1016/j.sajb.2010.01.002
dc.identifier.apacitationSmart, M., & Roden, L. C. (2010). A small-scale RNA isolation protocol useful for high-throughput extractions from recalcitrant plants. <i>South African Journal of Botany</i>, http://hdl.handle.net/11427/28044en_ZA
dc.identifier.chicagocitationSmart, Mariette, and Laura Catherine Roden "A small-scale RNA isolation protocol useful for high-throughput extractions from recalcitrant plants." <i>South African Journal of Botany</i> (2010) http://hdl.handle.net/11427/28044en_ZA
dc.identifier.citationSmart, M., & Roden, L. C. (2010). A small-scale RNA isolation protocol useful for high-throughput extractions from recalcitrant plants. South african journal of botany, 76(2), 375-379.
dc.identifier.ris TY - AU - Smart, Mariette AU - Roden, Laura Catherine AB - Many plants indigenous to South Africa are rich in secondary and oxidizing compounds such as pigments, complex polysaccharides and polyphenols. This makes isolation of high quality RNA for analysis of gene expression difficult. Here we describe a cost-effective isolation protocol suitable for RNA extraction from recalcitrant plant species. This method uses small amounts of tissue, so is useful when material is limited, and is easy to process large numbers of samples at once. We have used the method successfully with mature leaves of Protea hybrid ‘Sylvia’, and species P. repens, Leucospermum hybrid ‘Succession’, resurrection plants Xerophyta humilis and Craterostigma pumilum, and mature needles of Pine (Pinus radiata). RNA was analyzed spectrophotometrically and was found to be of high purity with low levels of contaminating compounds. Electrophoretic analyses on denaturing formaldehyde agarose gels and an Agilent 2100 Bioanalyzer confirmed the presence of RNA of high integrity. This is the first description of plant RNA integrity number (RIN) values for these plants using the algorithm designed for analyses of plant RNA containing multiple ribosomal bands. The RNA could successfully be used for reverse transcription and gene amplification. DA - 2010 DB - OpenUCT DP - University of Cape Town J1 - South African Journal of Botany LK - https://open.uct.ac.za PB - University of Cape Town PY - 2010 T1 - A small-scale RNA isolation protocol useful for high-throughput extractions from recalcitrant plants TI - A small-scale RNA isolation protocol useful for high-throughput extractions from recalcitrant plants UR - http://hdl.handle.net/11427/28044 ER - en_ZA
dc.identifier.urihttp://hdl.handle.net/11427/28044
dc.identifier.vancouvercitationSmart M, Roden LC. A small-scale RNA isolation protocol useful for high-throughput extractions from recalcitrant plants. South African Journal of Botany. 2010; http://hdl.handle.net/11427/28044.en_ZA
dc.language.isoeng
dc.publisher.departmentDepartment of Molecular and Cell Biologyen_ZA
dc.publisher.facultyFaculty of Scienceen_ZA
dc.publisher.institutionUniversity of Cape Town
dc.sourceSouth African Journal of Botany
dc.source.urihttps://www.journals.elsevier.com/south-african-journal-of-botany
dc.subject.otherRNA isolation
dc.subject.otherGene expression
dc.subject.otherRIN value
dc.subject.otherProteaceae
dc.subject.otherPine
dc.subject.otherResurrection plants
dc.titleA small-scale RNA isolation protocol useful for high-throughput extractions from recalcitrant plants
dc.typeJournal Article
uct.type.filetypeText
uct.type.filetypeImage
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