Simple and rapid protocol for the isolation of PCR-amplifiable DNA from medicinal plants

Abstract

ABSTRACT.Medicinal plant species has a valuable economic importance because of its usage as pharmaceuticals, nutritional, as well as its use in popular medication. For DNA-based techniques, nanogram quantities of the purified DNA are requisite to amplify and yield sufficient amounts of PCR products. SDS-based DNA isolation method was used to extract DNA from 11 species of different aromatic and medicinal plants collected from Saudi Arabia. Three hundred milligrams of fresh shredded plant material was necessary. The DNA purity was further confirmed by agarose gel, restriction endonuclease digestion and microsatellite primedpolymerase chain reaction (MP-PCR). DNA yields ranged from 10-20 µg (in 100-µL elution volumes) from all plant material evaluated. The DNA obtained was free of any contaminating proteins, polysaccharides and colored pigments. The extracted genomic DNA was found suitable for restriction digestion and PCR amplification. Our experimental procedure provides an easy, suitable, non-toxic, 349©FUNPEC-RP www.funpecrp.com.br Genetics and Molecular Research 11 (1): 348-354 (2012) Protocol for the isolation of PCR-amplifiable DNA cheap, and quick process for the amplification of DNA from medical plant tissue.

The Promise of Molecular and Genomic Techniques for Biodiversity Research and DNA Barcoding of the Arabian Peninsula Flora

“…These studies successfully demonstrated the use of SSR alleles to distinguish between large numbers of date palm cultivars. Alatar et al (2012) detected polymorphism in different medicinal plant species in Saudi Arabia and provided a protocol for DNA isolation and the SSR-PCR method. A microsatellite-based investigation of gene flow between wild barley Hordeum spontaneum and cultivated barley Hordeum vulgare was also performed by Hubner et al (2012) in Jordan.…”

A high-throughput, high-quality plant genomic DNA extraction protocol

“…Liquid nitrogen or dry ice is required for grinding in most traditional plant gDNA extraction methods (Dellaporta et al, 1983;Mohd-Hairul et al, 2011;Alatar et al, 2012;Mornkham et al, 2012;Souza et al, 2012). However, liquid nitrogen or dry ice cannot be obtained quickly in remote locations, especially in many developing countries.…”

“…Most conventional gDNA extraction methods are based on a 1.5-or 2.0-mL microcentrifuge tube format (Moslem et al, 2010;Mohd-Hairul et al, 2011;Alatar et al, 2012;Mornkham et al, 2012;Ojeda et al, 2012;Souza et al, 2012). Although it is effective in lowthroughput applications, these methods are inconvenient for a large number of samples.…”