Magnetic hypercrosslinked poly(styrene-co-divinylbenzene) microspheres (mgt.HPS1-NH2 and mgt.HPS2-NH2) containing different contents of amino groups were prepared and characterized in this study. The microspheres were used for the capture of uncompacted and compacted bacterial and calf thymus DNAs in the presence of different PEG 6000 and NaCl concentrations. Magnetic macroporous poly(glycidyl methacrylate-co-[2-(methacryloyloxy)ethoxy]acetic acid-co-ethylene dimethacrylate) microspheres containing amino and carboxyl groups [mgt.P(GMA-MOEAA-EDMA)-NH2] and magnetic non-porous carboxyl group-functionalized poly(2-hydroxyethyl methacrylate-co-glycidyl methacrylate) [mgt.P(HEMA-GMA)-COOH] microspheres were used as a control. The size changes of uncompacted and compacted bacterial and calf thymus DNAs were measured in the range of concentrations 0-16% PEG 6000 and 0.5-2.0 M NaCl by dynamic light scattering (DLS). The highest capture of compacted bacterial and calf thymus DNAs was achieved with mgt.HPS1-NH2, mgt.HPS2-NH2 microspheres using 8% PEG 6000 and 2.0 M NaCl and with mgt.P(GMA-MOEAA-EDMA)-NH2 microspheres using 8% and 16% PEG 6000 and 2.0 M NaCl. Developed microspheres were used for DNA isolation from real vegetable (broccoli flowering head) samples. DNA was in quality suitable for PCR.