The efficiency of solid phase extraction (SPE) of DNA on polymer particles is limited by the features of the applied solid support, such as size, hydrophilicity, and functionality and their application in SPE also requires additional steps and compounds to finally obtain sufficient amount of high-quality DNA. The present study describes a preparation of sub-micrometer monodisperse poly(methacrylic acid-co-ethylene dimethacrylate) (PME) particles by precipitation polymerization. The effect of the ethylene dimethacrylate (EDMA) crosslinker concentration on morphology and particle size, which varied from 730 to 900 nm, was investigated. The particles with 5 and 15 wt% EDMA were selected for a study of SPE of plasmid DNA under various adsorption and elution conditions, followed by the enzymatic restriction of isolated DNA to verify a quality the nucleic acid. The particles with 15 wt% EDMA were suitable for the SPE because they retained better colloidal stability during the adsorption without additional induction of DNA conformational change. The quality of isolated DNA was finally verified by enzymatic restriction by restriction endonuclease EcoRI. Moreover, the developed method using PME particles was successfully utilized for DNA isolation from Escherichia coli lysate.

• Klíčová slova
• DNA, EcoRI, Enzymatic restriction, Microparticles, Poly(methacrylic acid-co-ethylene dimethacrylate), Solid phase extraction,
• MeSH
• DNA bakterií chemie   izolace a purifikace   MeSH
• DNA chemie   izolace a purifikace   MeSH
• extrakce na pevné fázi * metody   MeSH
• hydrofobní a hydrofilní interakce MeSH
• koncentrace vodíkových iontů MeSH
• polymery chemie   MeSH
• polymethylmethakrylát chemie   MeSH
• velikost částic MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• DNA bakterií MeSH
• DNA MeSH
• poly(MAA-co-EDMA) MeSH Prohlížeč
• polymery MeSH
• polymethylmethakrylát MeSH

Cisplatin belongs to the most widely used cytostatic drugs. The determination of the presence of the DNA-cisplatin adducts may not only signal the guanine-rich regions but also monitor the interaction reaction between DNA and the drug in terms of speed of interaction. In this work, the combined advantages of magnetic particles-based isolation/purification with fluorescent properties of quantum dots (QDs) and antibodies targeted on specific recognition of DNA-cisplatin adducts are demonstrated. The formation of a complex between magnetic particles with surface modified by anti-dsDNA antibody, cisplatin-modified DNA and QDs labelled anti-cisplatin-modified DNA antibody was suggested and optimized.

• Klíčová slova
• Anti-DNA Antibodies, Cisplatin, Magnetic Separation, Sandwich Analysis,
• Publikační typ
• časopisecké články MeSH