Capture and release of DNA using aminosilane-modified bacterial magnetic particles for automated detection system of single nucleotide polymorphisms.
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.
Nakagawa
Takahito
T
Department of Biotechnology, Tokyo University of Agriculture and Technology, 2-24-16, Naka-cho, Koganei, Tokyo 184-8588, Japan.
Hashimoto
Reisuke
R
Maruyama
Kohei
K
Tanaka
Tsuyoshi
T
Takeyama
Haruko
H
Matsunaga
Tadashi
T
D001426:Bacterial Proteins / Q000737:chemistry*; D004252:DNA Mutational Analysis / Q000379:methods*; D006801:Humans; D018189:Immunomagnetic Separation / Q000379:methods*; D042201:Magnetospirillum / Q000737:chemistry*; D010316:Particle Size; D020641:Polymorphism, Single Nucleotide / Q000235:genetics*; D012821:Silanes / Q000737:chemistry; D014462:Ultrafiltration / Q000379:methods*
D001426:Bacterial Proteins; D012821:Silanes