reproducible rna preparation from sugarcane and citrus for functional genomic applications从甘蔗和柑橘可复制的rna制备功能性基因组的应用程序.pdfVIP

Hindawi Publishing Corporation International Journal of Plant Genomics Volume 2009, Article ID 765367, 13 pages doi:10.1155/2009/765367 Methodology Report Reproducible RNA Preparation from Sugarcane and Citrus for Functional Genomic Applications Mona B. Damaj,1 Phillip D. Beremand,2 Marco T. Buenrostro-Nava,1 Beth Riedel,2 Joe J. Molina,1 Siva P. Kumpatla,3 Terry L. Thomas,2 and T. Erik Mirkov1 1 Department of Plant Pathology and Microbiology, Texas AgriLife Research, Texas AM System, Weslaco, TX 78596, USA 2 Laboratory for Functional Genomics, Department of Biology, Texas AM University, College Station, TX 77843-3258, USA 3 Department of Trait Genetics and Technologies, Dow AgroSciences LLC, 9330 Zionsville Road, Indianapolis, IN 46268, USA Correspondence should be addressed to Terry L. Thomas, tlthomas@ and T. Erik Mirkov, e-mirkov@ Received 9 April 2009; Revised 12 September 2009; Accepted 13 October 2009 Recommended by Hongbin Zhang 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 divers