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

Authors :
Nakagawa T
Hashimoto R
Maruyama K
Tanaka T
Takeyama H
Matsunaga T
Source :
Biotechnology and bioengineering [Biotechnol Bioeng] 2006 Aug 05; Vol. 94 (5), pp. 862-8.
Publication Year :
2006

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 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 microg 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 dehydrogenase 2 (ALDH2) was demonstrated.<br /> ((c) 2006 Wiley Periodicals, Inc.)

Details

Language :
English
ISSN :
0006-3592
Volume :
94
Issue :
5
Database :
MEDLINE
Journal :
Biotechnology and bioengineering
Publication Type :
Academic Journal
Accession number :
16523525
Full Text :
https://doi.org/10.1002/bit.20904