Reproducible RNA Preparation from Sugarcane and Citrus for Functional Genomic Applications

• Published in: International Journal of Plant Genomics Vol. 2009; no. 1; pp. 146 - 158
• Main Authors: Damaj, Mona B., Beremand, Phillip D., Buenrostro-Nava, Marco T., Riedel, Beth, Molina, Joe J., Kumpatla, Siva P., Thomas, Terry L., Mirkov, T. Erik
• Format: Journal Article
• Language: English
• Published: United States Hindawi Limiteds 2009; Hindawi Publishing Corporation; Hindawi Limited
• Subjects: Chitinase; Citrus; Ethanol; Genomics; Infections; Metabolites; Methodology Report; Methods; Molecular weight; Polyphenols; Studies; Viral infections
• ISSN: 1687-5370; 1687-5389
• DOI: 10.1155/2009/765367

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.