A simple and rapid leaf genomic DNA extraction method for polymerase chain reaction analysis

Document Type: Brief Report

Authors

1 Department of Plant Biotechnology, National Institute of Genetic Engineering and Biotechnology (NIGEB), P.O. Box 14965/161, Tehran, I.R. Iran.

2 Faculty of Agriculture and plant breeding, University college of Agriculture and natural resources, Tehran university, P.O. Box 3158711167, Karaj, Iran.

Abstract

In plants, secondary metabolites and polysaccharides interfere with genomic isolation procedures and downstream reactions such as restriction enzyme analysis and gene amplification. The removal of such contaminants needs complicated and time-consuming protocols. In this study, a simple, rapid and efficient method for leaf DNA extraction was optimized. This method use small amount of plant material to reduce inhibitory agents (alkaloids, phenolic). The procedure involves homogenization of the plant leaf in extraction buffer, incubation at 60ºC, extraction by chloroform: iso-amyl alcohol and finally DNA precipitation by cold isopropanol. The results showed that the extracted DNA could be used directly for PCR.

Keywords