ABSTRACT
We describe the use of a hybrid magnetic nanocomposite (HMNC) for the extraction and purification of plasmid DNA (pDNA) from Escherichia coli aqueous solutions. The HMNC, which was synthesized via emulsion polymerization, was characterized by transmission electron microscopy, scanning electron microscopy, UV-Vis spectroscopy, Fourier transform infrared spectroscopy, dynamic light scattering and magnetic measurements. The results confirmed the incorporation of polyaniline (Pani) in its conducting form onto a core formed by the magnetic iron oxide, with the hybrid particles presenting an average size of (95⯱â¯30) nm and a saturation magnetization of 30 emu/g. The yield, purity and quality of the pDNA purified by using the Pani HMNC were evaluated by UV-Vis spectroscopy, agarose gel electrophoresis, and Polymerase Chain Reaction (PCR), respectively. An average yield of ~6.9⯵g was obtained in the DNA extraction process, with the collected material presenting a good purity (a â³260/280 ratio in the 1.68-1.82 range) and an excellent quality, as confirmed by subsequent PCR assays. Hence, this HMNC appears as a promising material for use in pDNA purification protocols, and we suggest that this novel HMNC-based methodology can be of general interest and find widespread application in different biomedical procedures.
Subject(s)
Aniline Compounds/chemistry , DNA/chemistry , Ferric Compounds/chemistry , Magnetics , Nanocomposites/chemistry , Plasmids , Silicon/chemistryABSTRACT
We examined the properties of the nanocomposite γ-Fe2O3@Chi@Pani as an adsorbent of deoxyribonucleic acid (DNA). As a model system, we used an aqueous solution of salmon sperm DNA, whose decreasing concentration was followed by monitoring the 260â¯nm UV-vis absorption. After adjusting the data collected to a Langmuir isotherm curve, we estimated the adsorption capacity (qe) of the nanocomposite as 49.5â¯mg/g. We also observed that the kinetic model of the DNA capture presents a mixed character, with both chemical mechanisms and intraparticle diffusion processes involved. When the MNC was used to extract the DNA from complex samples (human blood), a capture rate of 80â¯ng/µL was achieved, with the collected fraction exhibiting good quality, as evaluated by PCR analysis and electrophoresis assays. These results suggest that the γ-Fe2O3@Chi@Pani nanocomposite is a promising adsorbent for use in protocols for purification of DNA from complex samples.