Preliminary studies on a new way of nucleic acids extraction using hydrogel microsphere

Kamil K. Kamiński ,  Michał Szuwarzyński ,  Krzysztof Szczubiałka ,  Maria Nowakowska 

Jagiellonian University, Faculty of Chemistry, Ingardena 3, Kraków 30-060, Poland

Abstract

           In many areas of modern medicine and molecular biology the isolation of a nucleic acid from the biological material is a key process. It is a preliminary step for many procedures without which the study on DNA or RNA would not be possible [1]. Hence there is a need to develop better methods to perform these procedures. The most serious problem with the techniques used today is the use of toxic solvents are often harmful to the environment and the laboratory personnel performing isolation. This work presents the results of the studies on the use of polymeric hydrogels based on fully biocompatible and biodegradable biomaterials for this purpose.
          The process of binding nucleic acids is based on the attraction between the negatively charged DNA and positively charged surface of the hydrogel. A similar effect was observed for heparin [2]. Dextran used as a starting material was crosslinked to obtain hydrogel and functionalized with positively charged ammonium groups in the reaction with ammonium epoxide and diepoxide. In order to obtained hydrogel microspheres with dimensions of several microns the whole process was carried out in an inverted emulsion. The resulting material was investigated using optical microscopy and atomic force microscopy.


Figure 1. Atomic force microscope picture of the studied microspheres

The average size of the obtained structures, and size distribution was determined. The ability of microspheres to bind DNA and other anionic biopolymers was studied. Also, the the possibility of the interaction of the microspheres with bovine serum albumin was examined.
       A similar experiment was carried out for silica gel augmented with NH2 groups and quaternary ammonium groups. None of these silicone materials was able to decrease concentration of DNA or other polymers tested. This may suggest that in addition to the positive charge also the chemical structure of the adsorbing material used for DNA or RNA extraction is important.  

Acknowledgement: The project was operated within the Fundation for Polish Science Team Programme (TEAM/2008-2/6) and Ventures Programme (Ventures/2009-4/4) cofinanced by EU European Regional Development Fund

References:
[1] Biochemia, Berg J. M., Tymoczko J. L. Stryer L. PWN Warszawa 2005
[2] K. Kamiński, K. Zazakowny, K. Szczubiałka, M. Nowakowska, Biomacromolecules
     9, (11), s. 3129, 2008

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Presentation: Poster at VIII Multidyscyplinarna Konferencja Nauki o Leku, by Kamil K. Kamiński
See On-line Journal of VIII Multidyscyplinarna Konferencja Nauki o Leku

Submitted: 2012-02-22 23:54
Revised:   2012-02-23 00:01
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