Article
Evaluation of DNA extraction methods of rumen microbial populations
Registro en:
Villegas Rivera G., Vargas Cabrera Y., González Silva N., Aguilera García F., Gutiérrez Vázquez E., Bravo Patiño A., Cajero Juárez M., Baizabal Aguirre V.M., & Valdez Alarcón J.J.(2013). Evaluation of DNA extraction methods of rumen microbial populations. World J Microbiology Biotechnology (2013) 29:301–307. DOI 10.1007/s11274-012-1183-2
1573-0972
0959-3993
DOI 10.1007/s11274-012-1183-2
Autor
Villegas Rivera, Gabriela
Vargas Cabrera, Yevani
González Silva, Napoleón
Aguilera García, Florentino
Gutiérrez Vázquez, Ernestina
Bravo Patiño, Alejandro
Cajero Juárez, Marco
Baizabal Aguirre, Víctor Manuel
Valdez Alarcón, Juan José
Institución
Resumen
Artículo The dynamism of microbial populations in the rumen has been studied with molecular methods that analyze single nucleotide polymorphisms of ribosomal RNA gene fragments (rDNA). Therefore DNA of good quality is needed for this kind of analysis. In this work we report the evaluation of four DNA extraction protocols (mechanical lysis or chemical lysis with CTAB, ethylxanthogenate or DNAzol®) from ruminal fluid. The suitability of two of these protocols (mechanical lysis and DNAzol®) was tested on single-strand conformation polymorphism (SSCP) of rDNA of rumen microbial populations. DNAzol® was a simple method that rendered good integrity, yield and purity. With this method, subtle changes in protozoan populations were detected in young bulls fed with slightly different formulations of a supplement of multinutritional blocks of molasses and urea. Sequences related to Eudiplodinium maggi and a non-cultured Entodiniomorphid similar to Entodinium caudatum, were related to major fluctuating populations in an SSCP assay.