Capture and release of DNA using aminosilane-modified bacterial magnetic particles for automated detection system of single nucleotide polymorphisms
✍ Scribed by Takahito Nakagawa; Reisuke Hashimoto; Kohei Maruyama; Tsuyoshi Tanaka; Haruko Takeyama; Tadashi Matsunaga
- Publisher
- John Wiley and Sons
- Year
- 2006
- Tongue
- English
- Weight
- 265 KB
- Volume
- 94
- Category
- Article
- ISSN
- 0006-3592
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✦ Synopsis
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 Â 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 mg 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 808C 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.