Reproducible RNA Preparation from Sugarcane and Citrus for Functional Genomic Applications
Identifieur interne : 000B14 ( Ncbi/Curation ); précédent : 000B13; suivant : 000B15Reproducible RNA Preparation from Sugarcane and Citrus for Functional Genomic Applications
Auteurs : Mona B. Damaj [États-Unis] ; Phillip D. Beremand [États-Unis] ; Marco T. Buenrostro-Nava [États-Unis] ; Beth Riedel [États-Unis] ; Joe J. Molina [États-Unis] ; Siva P. Kumpatla [États-Unis] ; Terry L. Thomas [États-Unis] ; T. Erik Mirkov [États-Unis]Source :
- International Journal of Plant Genomics [ 1687-5370 ] ; 2009.
Abstract
High-throughput functional genomic procedures depend on the quality of the RNA used. Copurifying molecules can negatively impact the functionality of some plant RNA preparations employed in these procedures. We present a simplified, rapid, and scalable SDS/phenol-based method that provides the high-quantity and -quality RNA required by the newly emerging biotechnology applications. The method is applied to isolating RNA from tissues of two biotechnologically important crop plants, sugarcane and citrus, which provide a challenge due to the presence of fiber, polysaccharides, or secondary metabolites. The RNA isolated by this method is suitable for several downstream applications including northern blot hybridization, microarray analysis, and quantitative RT-PCR. This method has been used in a diverse range of projects ranging from screening plant lines overexpressing mammalian genes to analyzing plant responses to viral infection and defense signaling molecules.
Url:
DOI: 10.1155/2009/765367
PubMed: 20148085
PubMed Central: 2817868
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Kumpatla</name><affiliation wicri:level="1"><nlm:aff id="I3">Department of Trait Genetics and Technologies, Dow AgroSciences LLC, 9330 Zionsville Road, Indianapolis, IN 46268, USA</nlm:aff><country xml:lang="fr">États-Unis</country><wicri:regionArea>Department of Trait Genetics and Technologies, Dow AgroSciences LLC, 9330 Zionsville Road, Indianapolis, IN 46268</wicri:regionArea><wicri:noRegion>IN 46268</wicri:noRegion></affiliation></author><author><name sortKey="Thomas, Terry L" sort="Thomas, Terry L" uniqKey="Thomas T" first="Terry L." last="Thomas">Terry L. 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Erik Mirkov</name><affiliation wicri:level="1"><nlm:aff id="I1">Department of Plant Pathology and Microbiology, Texas AgriLife Research, Texas A&M System, Weslaco, TX 78596, USA</nlm:aff><country xml:lang="fr">États-Unis</country><wicri:regionArea>Department of Plant Pathology and Microbiology, Texas AgriLife Research, Texas A&M System, Weslaco, TX 78596</wicri:regionArea><wicri:noRegion>TX 78596</wicri:noRegion></affiliation></author></analytic><series><title level="j">International Journal of Plant Genomics</title><idno type="ISSN">1687-5370</idno><idno type="eISSN">1687-5389</idno><imprint><date when="2009">2009</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en"><p>High-throughput functional genomic procedures depend on the quality of the RNA used. Copurifying molecules can negatively impact the functionality of some plant RNA preparations employed in these procedures. We present a simplified, rapid, and scalable SDS/phenol-based method that provides the high-quantity and -quality RNA required by the newly emerging biotechnology applications. The method is applied to isolating RNA from tissues of two biotechnologically important crop plants, sugarcane and citrus, which provide a challenge due to the presence of fiber, polysaccharides, or secondary metabolites. The RNA isolated by this method is suitable for several downstream applications including northern blot hybridization, microarray analysis, and quantitative RT-PCR. This method has been used in a diverse range of projects ranging from screening plant lines overexpressing mammalian genes to analyzing plant responses to viral infection and defense signaling molecules.</p></div></front><back><div1 type="bibliography"><listBibl><biblStruct><analytic><author><name sortKey="Chaves, Mm" uniqKey="Chaves M">MM Chaves</name></author><author><name sortKey="Maroco, Jp" uniqKey="Maroco J">JP Maroco</name></author><author><name sortKey="Pereira, Js" uniqKey="Pereira J">JS Pereira</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Winkel Shirley, B" uniqKey="Winkel Shirley B">B Winkel-Shirley</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Salzman, Ra" uniqKey="Salzman R">RA Salzman</name></author><author><name sortKey="Fujita, T" uniqKey="Fujita T">T Fujita</name></author><author><name sortKey="Zhu Salzman, K" uniqKey="Zhu Salzman K">K Zhu-Salzman</name></author><author><name 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Lübberstedt</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Shi, C" uniqKey="Shi C">C Shi</name></author><author><name sortKey="Thummler, F" uniqKey="Thummler F">F Thümmler</name></author><author><name sortKey="Melchinger, Ae" uniqKey="Melchinger A">AE Melchinger</name></author><author><name sortKey="Wenzel, G" uniqKey="Wenzel G">G Wenzel</name></author><author><name sortKey="Lubberstedt, T" uniqKey="Lubberstedt T">T Lübberstedt</name></author></analytic></biblStruct></listBibl></div1></back></TEI></record>Pour manipuler ce document sous Unix (Dilib)
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