A rapid method for isolation of total DNA from pathogenic filamentous plant fungi
González-Mendoza, D; Argumedo-Delira, R; Morales-Trejo, A; Pulido-Herrera, A; Cervantes-Díaz, L; Grimaldo-Juarez, O; Alarcón, A
HERO ID
1940795
Reference Type
Journal Article
Year
2010
Language
English
PMID
| HERO ID | 1940795 |
|---|---|
| In Press | No |
| Year | 2010 |
| Title | A rapid method for isolation of total DNA from pathogenic filamentous plant fungi |
| Authors | González-Mendoza, D; Argumedo-Delira, R; Morales-Trejo, A; Pulido-Herrera, A; Cervantes-Díaz, L; Grimaldo-Juarez, O; Alarcón, A |
| Journal | Genetics and Molecular Research |
| Volume | 9 |
| Issue | 1 |
| Page Numbers | 162-166 |
| 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. |
| Doi | 10.4238/vol9-1gmr680 |
| Pmid | 20198572 |
| Wosid | WOS:000277326200011 |
| Is Certified Translation | No |
| Dupe Override | No |
| Is Public | Yes |
| Language Text | English |
| Keyword | Genomic DNA extraction; Fusarium oxysporum; Pyrenochaeta terrestris; Polymerase chain reaction; Filamentous fungi |