Krízová Jana, Spanová Alena, Rittich Bohuslav, Horák Daniel
Masaryk University Brno, Faculty of Science, Department of Microbiology, Tvrdého 14, 60200 Brno, Czech Republic.
J Chromatogr A. 2005 Feb 4;1064(2):247-53. doi: 10.1016/j.chroma.2004.12.014.
Carboxyl groups containing magnetic and non-magnetic microspheres were used in solid-phase reversible immobilization (SPRI) of genomic DNA. Magnetic non-porous poly(2-hydroxyethyl methacrylate-co-ethylene dimethacrylate)--P(HEMA-co-EDMA), poly(glycidyl methacrylate)--PGMA and P(HEMA-co-GMA) microspheres with hydrophilic properties were prepared by dispersion copolymerization of the respective monomers in the presence of colloidal iron oxides. DNA from chicken erythrocytes and DNA isolated from bacterial cells of Bifidobacterium longum was used for testing of adsorption/desorption properties of magnetic microspheres. The occurrence of false negative results in polymerase chain reaction (PCR) caused by the presence of extracellular inhibitors in DNA samples has been solved using SPRI. The P(HEMA-co-EDMA) and P(HEMA-co-GMA) microspheres were used for isolation of DNA from different dairy products followed by PCR identification of Bifidobacterium strains.
含羧基的磁性和非磁性微球用于基因组DNA的固相可逆固定化（SPRI）。通过在胶体氧化铁存在下使相应单体进行分散共聚，制备了具有亲水性的磁性无孔聚（甲基丙烯酸2-羟乙酯-共-二甲基丙烯酸乙二酯）——P（HEMA-co-EDMA）、聚（甲基丙烯酸缩水甘油酯）——PGMA和P（HEMA-co-GMA）微球。来自鸡红细胞的DNA和从长双歧杆菌细菌细胞中分离的DNA用于测试磁性微球的吸附/解吸特性。使用SPRI解决了DNA样品中细胞外抑制剂的存在导致聚合酶链反应（PCR）出现假阴性结果的问题。P（HEMA-co-EDMA）和P（HEMA-co-GMA）微球用于从不同乳制品中分离DNA，随后对双歧杆菌菌株进行PCR鉴定。
