Carboxyl-functionalized magnetic microparticle carrier for isolation and identification of DNA in Dairy products

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Authors

HORÁK Daniel RITTICH Bohuslav ŠPANOVÁ Alena

Year of publication 2006
Type Article in Proceedings
Conference 6th International Conference on the Scietific and Clinical applicationsof magnetic Carriers
MU Faculty or unit

Faculty of Science

Citation
Field Macromolecular chemistry
Keywords Magnetic particles; praparation; DNA separation; Lactobacillus; Bifidobacterium; PCR
Description Magnetite nanoparticles ca. 14 nm in diameter were obtained by chemical coprecipitation of Fe(II) and Fe(III) salts with ammonia in the presence of poly(ethylene glycol) (PEG). Magnetic poly(glycidyl methacrylate) (PGMA) microspheres were prepared by dispersion polymerization in poly(vinylpyrrolidone) (PVP)-stabilized aqueous ethanol with PEG-coated magnetite nanoparticles. Magnetic PGMA microspheres were hydrolyzed and carboxyl groups introduced by oxidation with KMnO4. Such a carrier was used for isolation of DNA. The particles reversibly bound DNA in the presence of high concentration of PEG 6,000 and sodium chloride. Bacterial DNA was isolated from crude cell lysates of different dairy products (butter milk, cheese, yoghurt), probiotic tablets or from cell lyophilisates. The presence of Bifidobacterium and Lactobacillus DNA in samples was identified by PCR amplification.
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