title: DNA recovery from a single bacterial cell using charge-reversible magnetic nanoparticles.
abstract: Highly efficient DNA recovery from a single bacterial cell was performed by means of imidazole-modified magnetic nanoparticles (Imi-MNPs). The modification by imidazole was confirmed by Fourier transform infrared spectroscopy. The Imi-MNPs were highly efficient at DNA extraction owing to the charge-reversible properties of Imi-MNPs, whereby DNA is attached to the particles at low pH and eluted at high pH because of electrostatic interactions. The DNA recovery ratio was determined by real-time PCR, and it revealed that complete recovery was guaranteed at ≥10(3) genome copies of Bacillus subtilis. Extraction of DNA from single bacterial cells was followed by PCR amplification of 16S rDNA and capillary electrophoresis. We achieved detection of single bacterial cells with a detection rate of 80%. We believe that our DNA recovery strategy may serve as a powerful tool for efficient DNA extraction and should be useful for quality control of cosmetics, foods, and pharmaceutical products.
authors.affiliation: Division of Biotechnology and Life Science, Institute of Engineering, Tokyo University of Agriculture and Technology, 2-24-16, Naka-cho, Koganei, Tokyo 184-8588, Japan.
authors.forename: Yoshiaki
authors.initials: Y
authors.lastname: Maeda
authors.affiliation: Division of Biotechnology and Life Science, Institute of Engineering, Tokyo University of Agriculture and Technology, 2-24-16, Naka-cho, Koganei, Tokyo 184-8588, Japan.
authors.forename: Takahiro
authors.initials: T
authors.lastname: Toyoda
authors.affiliation: Yokogawa Electric Corporation, 2-9-32, Naka-cho, Musashino-shi, Tokyo 180-8750, Japan.
authors.forename: Takeyuki
authors.initials: T
authors.lastname: Mogi
authors.affiliation: Yokogawa Electric Corporation, 2-9-32, Naka-cho, Musashino-shi, Tokyo 180-8750, Japan.
authors.forename: Tomoyuki
authors.initials: T
authors.lastname: Taguchi
authors.affiliation: Yokogawa Electric Corporation, 2-9-32, Naka-cho, Musashino-shi, Tokyo 180-8750, Japan.
authors.forename: Takeo
authors.initials: T
authors.lastname: Tanaami
authors.affiliation: Division of Biotechnology and Life Science, Institute of Engineering, Tokyo University of Agriculture and Technology, 2-24-16, Naka-cho, Koganei, Tokyo 184-8588, Japan; JST, CREST, Japan.
authors.forename: Tomoko
authors.initials: T
authors.lastname: Yoshino
authors.affiliation: Division of Biotechnology and Life Science, Institute of Engineering, Tokyo University of Agriculture and Technology, 2-24-16, Naka-cho, Koganei, Tokyo 184-8588, Japan.
authors.forename: Tadashi
authors.initials: T
authors.lastname: Matsunaga
authors.affiliation: Division of Biotechnology and Life Science, Institute of Engineering, Tokyo University of Agriculture and Technology, 2-24-16, Naka-cho, Koganei, Tokyo 184-8588, Japan; JST, CREST, Japan. Electronic address: tsuyo@cc.tuat.ac.jp.
authors.forename: Tsuyoshi
authors.initials: T
authors.lastname: Tanaka
mesh_terms: D001412:Bacillus subtilis / Q000737:chemistry*; D004269:DNA, Bacterial / Q000302:isolation & purification*; D019075:Electrophoresis, Capillary; D006863:Hydrogen-Ion Concentration; D007093:Imidazoles / Q000737:chemistry*; D058185:Magnetite Nanoparticles / Q000737:chemistry*; D012336:RNA, Ribosomal, 16S / Q000737:chemistry*; D060888:Real-Time Polymerase Chain Reaction; D059010:Single-Cell Analysis; D055672:Static Electricity
chemical_list: D004269:DNA, Bacterial; D007093:Imidazoles; D058185:Magnetite Nanoparticles; D012336:RNA, Ribosomal, 16S; C029899:imidazole
keywords: DNA recovery; Imidazole; Magnetic nanoparticles; Single bacterial cell