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Extraction of plasmid DNA by use of a magnetic maghemite-polyaniline nanocomposite

Author:
da Silva, Romário Justino, Maciel, Bruna Gomes, Medina-Llamas, Juan Carlos, Chávez-Guajardo, Alicia Elizabeth, Alcaraz-Espinoza, José Jarib, Pinto de Melo, Celso
Source:
Analytical biochemistry 2019 v.575 pp. 27-35
ISSN:
0003-2697
Subject:
Escherichia coli, Fourier transform infrared spectroscopy, agar gel electrophoresis, aqueous solutions, emulsions, light scattering, magnetism, nanocomposites, plasmids, polymerase chain reaction, polymerization, polymers, scanning electron microscopy, transmission electron microscopy, ultraviolet-visible spectroscopy
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 ± 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 μg was obtained in the DNA extraction process, with the collected material presenting a good purity (a ₳260/280 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.
Agid:
6350094