Marker analysis trans-gene detection and disease diagnosis by PCR analysis of a simple plant preparation

Many routine analyses of plants require the preparation or extraction of nucleic acid for PCR analysis. We have systematically evaluated the options for a single step protocol for the preparation of plant material. The objective was to establish the optimum composition of a solution in which a small sample of the plant tissue could be heated to prepare an extract that was suitable for direct introduction into a PCR. Heating tissue in 100 mM Tris-HCl, pH 7.4, 1 M KCl, 10 mM EDTA for 10 minutes at 95oC was found to give good results in a wide range of applications. The method was used for PCR analysis of single and multicopy genes, for RAPD analysis, for confirmation of transgenes and for analysis of DNA and RNA viruses in plant tissue. Samples successfully analysed included fresh, frozen and dried plant material, a wide range of plant species and plant tissues.