Enhanced solid phase extraction of DNA using hydrophilic monodisperse poly(methacrylic acid-co-ethylene dimethacrylate) microparticles
Jazyk angličtina Země Nizozemsko Médium print-electronic
Typ dokumentu časopisecké články
Grantová podpora
16-02702S
Grantová Agentura České Republiky
LO1507
Ministerstvo Školství, Mládeže a Tělovýchovy
PubMed
30859448
DOI
10.1007/s11033-019-04742-6
PII: 10.1007/s11033-019-04742-6
Knihovny.cz E-zdroje
- 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
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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