Isolation of Functional RNA from Heavily Infested, Wilted and Necrotic Leaf Tissues of Tea with High Polyphenol Content

A procedure is described to isolate intact RNA from tissues not previously undertaken-highly infested and wilted apical buds and leaves of tea (Camellia sinensis (L.) O. Ktze.). The protocol uses a final concentration of 450 mM β mercaptoethanol (βME) and 10% Polyvinylpyrrolidone (PVP) to circumvent problems associated with large amounts of polyphenols, polysaccharides, pigments and other secondary metabolites not easily removed by conventional procedures. Furthermore, the proposed protocol is applicable to normal tissues and other plant tissues with similar stresses, containing compounds that interfere with RNA extractions. Total RNA could be used for downstream applications such as mRNA isolation, reverse transcription, quantitative Polymerase Chain Reaction (PCR), cDNA library construction and Rapid amplification of cDNA ends (RACE).