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

被引:49
作者
Nakagawa, Takahito [1 ]
Hashimoto, Reisuke [1 ]
Maruyama, Kohei [1 ]
Tanaka, Tsuyoshi [1 ]
Takeyama, Haruko [1 ]
Matsunaga, Tadashi [1 ]
机构
[1] Tokyo Univ Agr & Technol, Dept Biotechnol, Tokyo 1848588, Japan
关键词
bacterial magnetic particles (BMPs); single nucleotide polymorphism (SNP); aminosilane compound; DNA capture; automated system; phosphate;
D O I
10.1002/bit.20904
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
Bacterial magnetic particles (BMPs) were modified with 3-[2-(2-aminoethylamino)-ethylaminol-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 x 10(4) 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 mu g 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 degrees 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 clehydrogenase 2 (ALDH2) was demonstrated. (c) 2006 Wiley Periodicals, Inc.
引用
收藏
页码:862 / 868
页数:7
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