Simplified extraction of good quality genomic DNA from a variety of plant materials

Abstract

Depending on the nature and complexity of plant material, proper method needs to be employed for extraction of genomic DNA, along with its performance evaluation by different molecular techniques. Here, we optimized and employed a simple genomic DNA isolation protocol suitable for a variety of plant materials covering in vitro grown tender plantlets to relatively complex plant tissues such as field grown mature potato leaves and tubers. Unlike other methods, no detergent was included in the isolation steps. This protocol, based on Dellaporta’s method as reported earlier, worked efficiently both at small and miniscale during handling large number of plant materials. DNA yield was found to be in the range of 70 to 120 μg per gram of the plant material; sufficient for most of the molecular techniques. Purity of DNA was checked by A₂₆₀/A₂₈₀ ratio, and restriction analyses including the isoschizomers HpaII and MspI. The DNA preparations were successfully used in polymerase chain reactions using genespecific primers for cloning of different genes. Prolonged storage did not affect the quality of the DNA samples. Taken together, this method could be a reliable substitute to frequently used chemical cetyl trimethylammonium bromide (CTAB) and commercial kits-based plant DNA isolation protocols.

Key words: Plant materials, genomic DNA isolation, restriction analyses, HpaII and MspI isoschizomers, genespecific primers, polymerase chain reaction (PCR), molecular cloning, DNA storage