Improved method for purification of bacterial DNA from bovine milk for detection of Brucella spp. by PCR

Different methods of extraction of bacterial DNA from bovine milk to improve the direct detection of Brucella by PCR were evaluated. We found that the use of a lysis buffer with high concentrations of Tris, EDTA, and NaCl, high concentrations of sodium dodecyl sulfate and proteinase K, and high temp...

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Bibliographic Details
Main Authors: Romero, C. (Conchi), Lopez-Goñi, I. (Ignacio)
Format: info:eu-repo/semantics/article
Language:eng
Published: American Society for Microbiology 2013
Subjects:
Online Access:https://hdl.handle.net/10171/29637
Description
Summary:Different methods of extraction of bacterial DNA from bovine milk to improve the direct detection of Brucella by PCR were evaluated. We found that the use of a lysis buffer with high concentrations of Tris, EDTA, and NaCl, high concentrations of sodium dodecyl sulfate and proteinase K, and high temperatures of incubation was necessary for the efficient extraction of Brucella DNA. The limit of detection by PCR was 5 to 50 Brucella CFU/ml of milk.