An effective method for isolating high quality RNA from spathe tissue of Anthurium andraeanum Hort. and other members of the Araceae family

Vern E. Collette, Paula E. Jameson, Kathy E. Schwinn, Chris S. Winefield, Pathmanathan Umaharan

Abstract


The abaxial and adaxial surfaces of Anthurium andraeanum Hort. (anthurium) spathe have marked cuticular thickening. In addition, the spathe tissue has high levels of polyphenols, evidenced by rapid browning upon injury. Both of these features hinder the extraction of high-quality RNA. To overcome these difficulties, an effective reproducible extraction method was developed using polyvinylpolypyrrolidone (PVPP) and 2?-mercaptoethanol. PVPP binds phenolics while 2?-mercaptoethanol prevents oxidation of phenolics. Both of these chemicals acting in concert possibly accounted for the marked improvement in the yield and quality of isolated RNA from anthurium spathe. Using this approach we obtained high quality RNA suitable for cDNA library construction, RT-PCR and northern analysis. The method was also effective with other members of the Araceae family.

Keywords


Anthurium, RNA extraction, spathe, Spathyphyllum, Zantedeschia

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