The E.Z.N.A^® Mag-Bind^® Plant DNA Kit is designed for the manual purification of DNA from plant leaves, roots and seed tissues. The system has been validated with cotton, sweet potato, soybean and tomato leaves. Up to 10 µg of high quality genomic DNA can be isolated from 50 gram of wet tissue (or 10 mg dry tissue) can be processed in less than 1 hour. This kit is designed for preparation of high quality genomic, chloroplast, and mitochondrial DNA. The DNA purified from these samples can be used in PCR as well as more demanding applications such as RAPD analysis.

Principle

The E.Z.N.A^® Mag-Bind^® Plant DNA Kit uses paramagnetic particles, considered a "mobile solid phase". The system combines the reversible nucleic acid-binding properties of Mag-Bind^® particles with the time-proven efficiency of SP plant lysis buffer system to eliminate polysaccharides, phenolic compounds, and enzyme inhibitors from plant tissue lysates. Unlike column-based systems, the binding of nucleic acids to magnetic particles occurs in solution, resulting in increased binding kinetics and binding efficiency. Particles are also completely re-suspended during the wash steps of the purification protocol, thus enhancing contact with the wash buffer and removal of contaminants and increasing nucleic acid purity.

Procedure

Dry or fresh plant tissue is disrupted and then lysed in a specially formulated buffer. Proteins, polysaccharides, and cellular debris are subsequently precipitated. The lysate is transferred to a new tube and binding conditions are adjusted so that genomic DNA will selectively bind to the MagBead^®. Two rapid wash steps remove trace contaminants such as residual polysaccharides, and pure DNA is eluted in Elution Buffer. Purified DNA can be directly used in downstream applications without the need for further purification.

Applications

DNA purified using the E.Z.N.A.^® Mag-Bind^® Plant DNA Kit is suitable for the following applications :

• PCR
• Restriction Digestion
• Hybridization Techniques
• RAPD analysis