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Enhanced solid phase extraction of DNA using hydrophilic monodisperse poly(methacrylic acid-co-ethylene dimethacrylate) microparticles

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Abstract

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.

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Acknowledgements

This work was supported by the Czech Science Foundation (No. 16-02702S) and the Ministry of Education, Youth and Sports (POLYMAT #LO1507).

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  1. Petr Šálek and Marcela Filipová have contributed equally to this work.

Authors and Affiliations

  1. Institute of Macromolecular Chemistry, Academy of Sciences of the Czech Republic, Heyrovského nám. 2, 162 06, Prague 6, Czech Republic

    Petr Šálek, Marcela Filipová, Daniel Horák, Vladimír Proks & Olga Janoušková

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  1. Petr Šálek
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Correspondence to Petr Šálek.

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Šálek, P., Filipová, M., Horák, D. et al. Enhanced solid phase extraction of DNA using hydrophilic monodisperse poly(methacrylic acid-co-ethylene dimethacrylate) microparticles. Mol Biol Rep 46, 3063–3072 (2019). https://doi.org/10.1007/s11033-019-04742-6

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