Isolation of microbial DNA by newly designed magnetic particles.
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) from dairy products, lyophilised cell cultures, and bacterial colonies grown on hard media, and Trichophyton fungi DNA from lyophilised cells. 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 cobalt ferrite nanoparticles modified with alginic acid and checked by restriction analysis.
Rittich
B
B
Masaryk University, Faculty of Science, CZ-611 37 Brno, Czech Republic. rittich@sci.muni.cz
Spanová
A
A
Horák
D
D
Benes
M J
MJ
Klesnilová
L
L
Petrová
K
K
Rybnikár
A
A
D001644:Bifidobacterium / Q000235:genetics / Q000302:isolation & purification; D004269:DNA, Bacterial / Q000235:genetics / Q000302:isolation & purification*; D004271:DNA, Fungal / Q000235:genetics / Q000302:isolation & purification*; D003611:Dairy Products / Q000382:microbiology; D005290:Ferric Compounds; D007779:Lactobacillus acidophilus / Q000235:genetics / Q000302:isolation & purification; D008280:Magnetics; D008855:Microscopy, Electron, Scanning; D008863:Microspheres; D053758:Nanoparticles; D011073:Polyamines; D011102:Polyhydroxyethyl Methacrylate / Q000031:analogs & derivatives; D016133:Polymerase Chain Reaction; D011109:Polymethacrylic Acids; D014249:Trichophyton / Q000235:genetics / Q000302:isolation & purification
D004269:DNA, Bacterial; D004271:DNA, Fungal; D005290:Ferric Compounds; D011073:Polyamines; D011109:Polymethacrylic Acids; C056971:poly(2-hydroxyethyl methacrylate)-polyamine graft copolymer; C001215:ferrite; C042535:polyglycidyl methacrylate; D011102:Polyhydroxyethyl Methacrylate