Methodology article

A fully automatable enzymatic method for DNA extraction from plant tissues

Jean-François Manen^{* 1} , Olga Sinitsyna^{* 2} , Lorène Aeschbach¹ , Alexander V Markov² and Arkady Sinitsyn^{* 2}

¹University of Geneva, Conservatoire et Jardin Botaniques de la Ville de Genève, Impératrice 1, CH-1292 Chambésy/Genève, Switzerland

²Moscow State University, Chemistry Department, Vorobyevy Gory, 119899, Moscow, Russia

author email corresponding author email* Contributed equally

BMC Plant Biology 2005, 5:23doi:10.1186/1471-2229-5-23

Published:	3 November 2005

Abstract

Background

DNA extraction from plant tissues, unlike DNA isolation from mammalian tissues, remains difficult due to the presence of a rigid cell wall around the plant cells. Currently used methods inevitably require a laborious mechanical grinding step, necessary to disrupt the cell wall for the release of DNA.

Results

Using a cocktail of different carbohydrases, a method was developed that enables a complete digestion of the plant cell walls and subsequent DNA release. Optimized conditions for the digestion reaction minimize DNA shearing and digestion, and maximize DNA release from the plant cell. The method gave good results in 125 of the 156 tested species.

Conclusion

In combination with conventional DNA isolation techniques, the new enzymatic method allows to obtain high-yield, high-molecular weight DNA, which can be used for many applications, including genome characterization by AFLP, RAPD and SSR. Automation of the protocol (from leaf disks to DNA) is possible with existing workstations.