Abstract

The isolation of un-degraded, high-molecular-mass genomic DNA is essential for many DNA based molecular biology applications including long polymerase chain reactions (PCR), multiplex PCR, endonuclease digestion, Southern blot analysis and genomic DNA library construction studies. Although many protocols are available for the extraction of DNA from plant species, most of them require longer times and the size of extracted DNA is usually below 20 kb. Most of commercially available kits are expensive, unavailable in many part of the world and also require at least 1.5 h including the RNase digestion. In the present study, we report a DNA extraction method using the MAGi reagent. The MAGi DNA extraction method extracts genomic DNAs with excellent spectral readings, efficiently digestible with endonucleases and suitable for enzymatic and multiplex PCR amplification studies. The MAGi DNA extraction method is suitable for multiplex PCR, endonuclease digestion studies and greatly reduces the time required to isolate high intact genomic DNAs from fresh leaf, fruit and fresh and dry seed at different development stages. Using the MAGi extraction protocol, highly pure DNA higher than 50 kb DNA in size can be obtained in approximately 1 h.

Key words: High quality DNA, MAGi solution, marker assisted selection (MAS), multiplex polymerase chain reaction (PCR).