Isolation of microbial DNA by newly designed magnetic particles

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Authors

RITTICH Bohuslav ŠPANOVÁ Alena HORÁK Daniel BENEŠ Milan KLESNILOVÁ Lenka PETROVÁ Kateřina RYBNIKÁŘ Alois

Year of publication 2006
Type Article in Periodical
Magazine / Source Colloids and Surfaces B: Biointerferences
MU Faculty or unit

Faculty of Science

Citation
Field Microbiology, virology
Keywords Magnetic particles P(HEMA-co-GMA); cobalt ferrite magnetic paricles; microbial DNA; polymerase chain reaction (PCR)
Description Carboxyl group-containing magnetic nonporous poly(2-hydroxyethyl methacrylate-co-glycidyl methacrylate) (P(HEMA-co-GMA)) microspheres and cobalt ferrite nanoparticles modified with alginic acid (natural carboxylic polysaccharide) were used for isolation of microbial DNA of lactic acid bacteria (LAB) and Trichophyton fungi DNA from lyophilized samples, from a bacterial colony grown on hard media, and from dairy products. DNA from the samples with lysed cells was reversibly adsorbed to the particles in the presence of high poly(ethylene glycol) (PEG 6000) and sodium chloride concentrations. The optimal final PEG and NaCl concentrations were 9.1 wt. % and 2.0 M, respectively. The adsorbed DNA was released from the particles in low ionic strength TE buffer. The quality of isolated DNA was checked by PCR amplification. Moreover, PCR amplicons were isolated on P(HEMA-co-GMA) microspheres and checked by restriction analysis.
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