A simple and efficient protocol for isolation of functional RNA from plant tissues rich in secondary metabolites

A protocol is described for rapid RNA isolation from various plant species and tissues rich in polyphenolics and polysaccharides. The method is based on the Nucleon PhytoPure(TM) system without the use of phenol. The procedure can be completed withi 1 h and many samples can be processed at the same time. The yield ranged from 240 mu g up to 3 mg per gram of tissue with an average purity measured as A(260/280) Of 1.85 The RNA was of sufficient quality fur use in RT-PCR reactions. Quantitation of single-stranded cDNA was carried out with the RiboGreen(TM) reagent and of PCR products with the PicoGreen(TM) reagent.