Capture and release of DNA using aminosilane-modified bacterial magnetic particles for automated detection system of single nucleotide polymorphisms

Takahito Nakagawa, Reisuke Hashimoto, Kohei Maruyama, Tsuyoshi Tanaka, Haruko Takeyama, Tadashi Matsunaga

研究成果: Article

43 引用 (Scopus)

抄録

Bacterial magnetic particles (BMPs) were modified with 3-[2-(2- aminoethylamino)-ethylamino]-propyltrimethoxysilane (AEEA) to produce a dense amine surface. Modification of BMPs in a toluene solution resulted in an increased amine yield, and approximately 11.3 × 104 surface amines were detected on a single particle. The modified BMPs were capable of efficient electrostatic capture of DNA. The maximum amount of DNA captured on 10 μg of aminosilane-modified BMPs was 600 ng. A 10 mM phosphate buffer effectively released the captured DNA. This efficiency was dramatically enhanced by incubation at 80°C and DNA recovery from aminosilane-modified BMPs approached 95%. DNA extraction from whole blood using these modified BMPs, followed by PCR, was successfully performed. Furthermore, automated single nucleotide polymorphism (SNP) detection of the aldehyde dehydrogenase 2 (ALDH2) was demonstrated.

元の言語English
ページ(範囲)862-868
ページ数7
ジャーナルBiotechnology and Bioengineering
94
発行部数5
DOI
出版物ステータスPublished - 2006 8 5
外部発表Yes

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Nucleotides
Polymorphism
Single Nucleotide Polymorphism
DNA
Amines
Aldehyde Dehydrogenase
Toluene
Aldehydes
Static Electricity
Electrostatics
Buffers
Phosphates
Blood
Recovery
Polymerase Chain Reaction

ASJC Scopus subject areas

  • Biotechnology
  • Microbiology

これを引用

Capture and release of DNA using aminosilane-modified bacterial magnetic particles for automated detection system of single nucleotide polymorphisms. / Nakagawa, Takahito; Hashimoto, Reisuke; Maruyama, Kohei; Tanaka, Tsuyoshi; Takeyama, Haruko; Matsunaga, Tadashi.

:: Biotechnology and Bioengineering, 巻 94, 番号 5, 05.08.2006, p. 862-868.

研究成果: Article

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abstract = "Bacterial magnetic particles (BMPs) were modified with 3-[2-(2- aminoethylamino)-ethylamino]-propyltrimethoxysilane (AEEA) to produce a dense amine surface. Modification of BMPs in a toluene solution resulted in an increased amine yield, and approximately 11.3 × 104 surface amines were detected on a single particle. The modified BMPs were capable of efficient electrostatic capture of DNA. The maximum amount of DNA captured on 10 μg of aminosilane-modified BMPs was 600 ng. A 10 mM phosphate buffer effectively released the captured DNA. This efficiency was dramatically enhanced by incubation at 80°C and DNA recovery from aminosilane-modified BMPs approached 95{\%}. DNA extraction from whole blood using these modified BMPs, followed by PCR, was successfully performed. Furthermore, automated single nucleotide polymorphism (SNP) detection of the aldehyde dehydrogenase 2 (ALDH2) was demonstrated.",
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AU - Nakagawa, Takahito

AU - Hashimoto, Reisuke

AU - Maruyama, Kohei

AU - Tanaka, Tsuyoshi

AU - Takeyama, Haruko

AU - Matsunaga, Tadashi

PY - 2006/8/5

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N2 - Bacterial magnetic particles (BMPs) were modified with 3-[2-(2- aminoethylamino)-ethylamino]-propyltrimethoxysilane (AEEA) to produce a dense amine surface. Modification of BMPs in a toluene solution resulted in an increased amine yield, and approximately 11.3 × 104 surface amines were detected on a single particle. The modified BMPs were capable of efficient electrostatic capture of DNA. The maximum amount of DNA captured on 10 μg of aminosilane-modified BMPs was 600 ng. A 10 mM phosphate buffer effectively released the captured DNA. This efficiency was dramatically enhanced by incubation at 80°C and DNA recovery from aminosilane-modified BMPs approached 95%. DNA extraction from whole blood using these modified BMPs, followed by PCR, was successfully performed. Furthermore, automated single nucleotide polymorphism (SNP) detection of the aldehyde dehydrogenase 2 (ALDH2) was demonstrated.

AB - Bacterial magnetic particles (BMPs) were modified with 3-[2-(2- aminoethylamino)-ethylamino]-propyltrimethoxysilane (AEEA) to produce a dense amine surface. Modification of BMPs in a toluene solution resulted in an increased amine yield, and approximately 11.3 × 104 surface amines were detected on a single particle. The modified BMPs were capable of efficient electrostatic capture of DNA. The maximum amount of DNA captured on 10 μg of aminosilane-modified BMPs was 600 ng. A 10 mM phosphate buffer effectively released the captured DNA. This efficiency was dramatically enhanced by incubation at 80°C and DNA recovery from aminosilane-modified BMPs approached 95%. DNA extraction from whole blood using these modified BMPs, followed by PCR, was successfully performed. Furthermore, automated single nucleotide polymorphism (SNP) detection of the aldehyde dehydrogenase 2 (ALDH2) was demonstrated.

KW - Aminosilane compound

KW - Automated system

KW - Bacterial magnetic particles (BMPs)

KW - DNA capture

KW - Phosphate

KW - Single nucleotide polymorphism (SNP)

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