A rapid and inexpensive method for isolation of total  DNA from Trichoderma spp (Hypocreaceae).
Extraction of high-quality genomic DNA for PCR amplification  from filamentous fungi is difficult because of the complex cell wall and the  high concentrations of polysaccharides and other secondary metabolites that  bind to or co-precipitate with nucleic acids. We developed a modified sodium  dodecyl sulfate/phenol protocol, without maceration in liquid nitrogen and  without a final ethanol precipitation step. The A(260/280) absorbance ratios of isolated DNA were approximately  1.7-1.9, demonstrating that the DNA fraction is pure and can be used for  analysis. Additionally, the A(260/230) values  were higher than 1.6, demonstrating 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. The main advantages of  the method are that the mycelium is directly recovered from culture medium and  it does not require the use of expensive and specialized equipment.
Instituto de Ciencias Agrícolas, Universidad Autónoma de Baja California, Baja California, México.
J C
JC
Vazquez-Angulo
V
V
Mendez-Trujillo
D
D
González-Mendoza
A
A
Morales-Trejo
O
O
Grimaldo-Juarez
L
L
Cervantes-Díaz
D004271:DNA, Fungal / Q000235:genetics*; D005658:Fungi / Q000235:genetics; D016133:Polymerase Chain Reaction; D014242:Trichoderma / Q000235:genetics*
D004271:DNA, Fungal