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Capture and release of DNA using aminosilane‐modified bacterial magnetic particles for automated detection system of single nucleotide polymorphisms
Author(s) -
Nakagawa Takahito,
Hashimoto Reisuke,
Maruyama Kohei,
Tanaka Tsuyoshi,
Takeyama Haruko,
Matsunaga Tadashi
Publication year - 2006
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/bit.20904
Subject(s) - dna , amine gas treating , chemistry , chromatography , nucleotide , dna extraction , single nucleotide polymorphism , toluene , microbiology and biotechnology , biochemistry , polymerase chain reaction , analytical chemistry (journal) , biology , gene , organic chemistry , genotype
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 × 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 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. © 2006 Wiley Periodicals, Inc.