One-Tube Preparation and PCR Amplification of DNA From Plant Leaf Tissue with Extract-N-AmpTM

The use of PCR amplification to detect target DNA sequences has many applications in plant genotyping, gene mapping, diagnostics, and diversity assessment. PCR itself is simple to set up, and requires little hands-on time. However, most methods for preparing DNA from plant tissues are time consuming, tedious, and labor intensive. Virtually all require mechanical disruption, such as grinding in liquid nitrogen1 or reciprocal shaking2, to break the plant cell wall. Furthermore, many use multiple extraction steps with organic solvents (phenol, chloroform), detergents (CTAB, SDS)1, salts (NaCl, ammonium acetate),3 and/or polyvinylpyrrollidone (PVP)3 to remove polysaccharides and polyphenolic components that can inhibit enzymatic reactions. Clearly, none of these methods is amenable to rapid analysis with more than a few test samples.