TY - JOUR
T1 - Extraction of plasmid DNA by use of a magnetic maghemite-polyaniline nanocomposite
AU - da Silva, Romário Justino
AU - Maciel, Bruna Gomes
AU - Medina-Llamas, Juan Carlos
AU - Chávez-Guajardo, Alicia Elizabeth
AU - Alcaraz-Espinoza, José Jarib
AU - Pinto de Melo, Celso
N1 - Publisher Copyright:
© 2019
PY - 2019/6/15
Y1 - 2019/6/15
N2 - 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.
AB - 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.
KW - Conducting polymers
KW - Escherichia coli
KW - Magnetic nanocomposite
KW - Magnetic purification
KW - Plasmid DNA
UR - http://www.scopus.com/inward/record.url?scp=85063632771&partnerID=8YFLogxK
U2 - 10.1016/j.ab.2019.03.013
DO - 10.1016/j.ab.2019.03.013
M3 - Artículo
C2 - 30917944
AN - SCOPUS:85063632771
SN - 0003-2697
VL - 575
SP - 27
EP - 35
JO - Analytical Biochemistry
JF - Analytical Biochemistry
ER -