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

Research output: Contribution to journalArticle

44 Citations (Scopus)

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.

Original languageEnglish
Pages (from-to)862-868
Number of pages7
JournalBiotechnology and Bioengineering
Volume94
Issue number5
DOIs
Publication statusPublished - 2006 Aug 5
Externally publishedYes

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Keywords

  • Aminosilane compound
  • Automated system
  • Bacterial magnetic particles (BMPs)
  • DNA capture
  • Phosphate
  • Single nucleotide polymorphism (SNP)

ASJC Scopus subject areas

  • Biotechnology
  • Microbiology

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