An Improved and Rapid Protocol for the Isolation of Polysaccharide- and Polyphenol-Free Sugarcane DNA

Abstract

We have optimized a simple and rapid method for isolating milligram quantities of high quality DNA from polysaccharide- and polyphenolic-rich tissue such as sugarcane, lettuce and strawberry. The protocol utilizes fresh tissue without making use of liquid nitrogen or freeze-drying for initial grinding of the tissue and it significantly minimize the use of lab materials. At least one hundred samples can be processed daily by one person. The isolated DNA is essentially free of polysaccharides, polyphenols, RNA and other major contaminants, as judged by: its clear color, its viscosity, A260/A280 ratio, digestibility with restriction enzymes, and suitability for Restriction Fragment Length Polymorphism (RFLP)- and Polymerase Chain Reaction (PCR)-based techniques.