DNA recovery from a single bacterial cell using charge-reversible magnetic nanoparticles

Yoshiaki Maeda; Takahiro Toyoda; Takeyuki Mogi; Tomoyuki Taguchi; Takeo Tanaami; Tomoko Yoshino; Tadashi Matsunaga; Tsuyoshi Tanaka

doi:10.1016/j.colsurfb.2015.11.057

DNA recovery from a single bacterial cell using charge-reversible magnetic nanoparticles

Yoshiaki Maeda, Takahiro Toyoda, Takeyuki Mogi, Tomoyuki Taguchi, Takeo Tanaami, Tomoko Yoshino, Tadashi Matsunaga, Tsuyoshi Tanaka^*

^*Corresponding author for this work

Research output: Contribution to journal › Article › peer-review

9 Citations (Scopus)

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 ≥10³ 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.

Original language	English
Pages (from-to)	117-122
Number of pages	6
Journal	Colloids and Surfaces B: Biointerfaces
Volume	139
DOIs	https://doi.org/10.1016/j.colsurfb.2015.11.057
Publication status	Published - 2016 Mar 1
Externally published	Yes

Keywords

DNA recovery
Imidazole
Magnetic nanoparticles
Single bacterial cell

ASJC Scopus subject areas

Biotechnology
Surfaces and Interfaces
Physical and Theoretical Chemistry
Colloid and Surface Chemistry

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10.1016/j.colsurfb.2015.11.057

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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 ≥103 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.",

keywords = "DNA recovery, Imidazole, Magnetic nanoparticles, Single bacterial cell",

author = "Yoshiaki Maeda and Takahiro Toyoda and Takeyuki Mogi and Tomoyuki Taguchi and Takeo Tanaami and Tomoko Yoshino and Tadashi Matsunaga and Tsuyoshi Tanaka",

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doi = "10.1016/j.colsurfb.2015.11.057",

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AU - Maeda, Yoshiaki

AU - Toyoda, Takahiro

AU - Mogi, Takeyuki

AU - Taguchi, Tomoyuki

AU - Tanaami, Takeo

AU - Yoshino, Tomoko

AU - Matsunaga, Tadashi

AU - Tanaka, Tsuyoshi

PY - 2016/3/1

Y1 - 2016/3/1

N2 - 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 ≥103 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.

AB - 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 ≥103 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.

KW - DNA recovery

KW - Imidazole

KW - Magnetic nanoparticles

KW - Single bacterial cell

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JO - Colloids and Surfaces B: Biointerfaces

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ER -

DNA recovery from a single bacterial cell using charge-reversible magnetic nanoparticles

Abstract

Keywords

ASJC Scopus subject areas

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