The Omega Biotek E-Z 96 Plant DNA DS Kit (Difficult Sample) is designed for efficient recovery of genomic DNA up to 30 kb in size from fresh, frozen, or dried plant tissue samples rich in polysaccharides, polyphenols, or those having a lower DNA content.  Up to 50 mg wet tissue can be processed in less than 1 hour.  The system combines the reversible nucleic acid-binding properties of the HiBind® matrix with the speed and versatility of spin column technology to eliminate polysaccharides, phenolic compounds, and enzyme inhibitors from plant tissue lysates.  Purified DNA is suitable for PCR, restriction digestion, and hybridization applications.

This procedure relies on the well-established properties of the cationic detergent, cetyltrimethyl ammonium bromide (CTAB), in conjunction with the unique binding system to increase yields and provide high-quality DNA.  The system eliminates the need for chloroform extractions traditionally associated with with CTAB-based lysis methods.  Samples are homogenized and lysed in a high salt buffer containing CTAB, binding conditions are adjusted, and DNA is purified using a E-Z 96 DNA Plate.  Salts, proteins, and other contaminants are removed to yield high-quality genomic DNA suitable for downstream applications such as endonuclease digestion, thermal cycle amplification, and hybridization applications.

Comparison of DNA yield from multiple crops. 40-50 mg of respective fresh leaf tissue was extracted in triplicate according to manufacturer’s recommended protocols and eluted in 100 µL. DNA analyzed with fluorescent DNA-based quantification method. Total yield was divided by total tissue amount to show ng of DNA per mg of leaf tissue.