Format

Send to

Choose Destination
Genet Mol Res. 2010 Feb 2;9(1):162-6. doi: 10.4238/vol9-1gmr680.

A rapid method for isolation of total DNA from pathogenic filamentous plant fungi.

Author information

1
Instituto de Ciencias Agrícolas, Universidad Autónoma de Baja California, Ejido Nuevo León, Baja California, México. daniasaf@gmail.com

Abstract

DNA isolation from some fungal organisms of agronomic importance is difficult because they have cell walls or capsules that are relatively unsusceptible to lysis. We have developed a fast DNA isolation protocol for Fusarium oxysporum, which causes fusarium wilt disease in more than 100 plant species, and for Pyrenochaeta terrestris, which causes pink root in onions. This protocol was based on the sodium dodecyl sulfate/phenol method, without beta-mercaptoethanol and without maceration in liquid nitrogen; it uses phenol/chloroform extraction to remove proteins and co-precipitated polysaccharides. The A(260/280) absorbance ratios of isolated DNA were around 1.9, suggesting that the DNA fraction was pure and may be used for further analysis. Additionally, the A(260/230) values were higher than 1.8, suggesting negligible contamination by polysaccharides. The DNA isolated by this protocol is of sufficient quality for molecular applications; this technique could be applied to other organisms that have similar substances that hinder DNA extraction.

PMID:
20198572
DOI:
10.4238/vol9-1gmr680
[Indexed for MEDLINE]
Free full text

Supplemental Content

Full text links

Icon for Fundacao de Pesquisas Cientificas de Ribeirao Preto
Loading ...
Support Center