Publication detail

Comparison and optimalization of DNA extraction methods using new magnetic particles for analysis of Haloarcula

KONEČNÁ, J. HORÁK, D. ŠPANOVÁ, A.

Original Title

Comparison and optimalization of DNA extraction methods using new magnetic particles for analysis of Haloarcula

Type

conference proceedings

Language

English

Original Abstract

Haloarcula used in this study was previously isolated from salt evaporation ponds belonging to Salinas del Odiel S.L. Company, located in the natural reserve of Odiel marshlands in the Southwest Spain1. DNA extraction is an essential step in all molecular biologist diagnostic methods. We compare three used methods, and found their DNA was changed in quality and quantity. A fundamental challenge in using these approaches has been to isolate DNA that is representative of the Haloarcula culture. Three different DNA extraction, performed with commercial kit (GeneJET Genomic DNA Purification Kit. Thermo Scientific, Wilmington, USA), a standard phenol-chloroform method and two magnetic micro- and nanoparticles were evaluated, using the addition of Haloarcula cells. DNA extracts were analysed by agarose gel electrophoresis and UV spectroscopy. Two different types of magnetic particles, prepared by Horák in Institute of Macromolecular Chemistry AS CR, were used for solid - phase DNA extraction. First types of magnetic microparticles was P(HEMA-GMA). P(HEMA) exhibits low protein adsorption and less cell adhesion. In attempt to promote cell adhesion, several methods were used to modify P(HEMA). Polymer modification have been shown to have a potential in enhancing the ability of a material to support cell adhesion2. Second studied magnetic nanoparticles was F79/L3 poly-LYSINE. In attempt to increase cell adhesion, the iron oxide surface was modified with poly-LYSINE (PLL). Positively charged interface clearly possessing an affinity for negatively charged cells. PLL is known to adsorb proteins not only from nutrient solutions, but also from cell membranes3. In the presented work was obtained intactness and suitable DNA by using magnetic particles for another sensitivity molecular-diagnostic methods.

Keywords

Haloarcula, magnetic nano- and microparticles, extraction methods

Authors

KONEČNÁ, J.; HORÁK, D.; ŠPANOVÁ, A.

Released

18. 6. 2017

Pages from

255

Pages to

255

Pages count

1

BibTex

@proceedings{BUT138053,
  editor="Jana {Konečná} and Daniel {Horák} and Alena {Španová}",
  title="Comparison and optimalization of DNA extraction methods using new magnetic particles for analysis of Haloarcula",
  year="2017",
  pages="255--255"
}