A fully automatable enzymatic method for DNA extraction from plant tissues.
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.


CONCLUSIONS
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.
Manen
Jean-François
JF
University of Geneva, Conservatoire et Jardin Botaniques de la Ville de Genève, Impératrice 1, CH-1292 Chambésy/Genève, Switzerland. manen@cjb.ville-ge.ch
Sinitsyna
Olga
O
Aeschbach
Lorène
L
Markov
Alexander V
AV
Sinitsyn
Arkady
A
D001901:Botany / Q000379:methods; D002473:Cell Wall / Q000378:metabolism; D018744:DNA, Plant / Q000302:isolation & purification*; D005821:Genetic Techniques; D018515:Plant Leaves / Q000235:genetics; D014242:Trichoderma / Q000201:enzymology*
D018744:DNA, Plant