DNA extraction using modified bacterial magnetic particles in the presence of amino silane compound.
Magnetic particles produced by magnetic bacteria have been used to carry out magnetic separation of DNA. Separation was achieved using magnetite coated with 3-aminopropyltriethoxysilane, N-(trimethoxy-silylpropyl) isothiouronium chloride or 3-[2-(2-aminoethyl)-ethylamino]-propyltrimethoxysilane (AEEA). The DNA binding efficiency increased with the number of amino groups present on the silane compounds and was 14 fold higher than with untreated magnetite. Addition of AEEA to aqueous solutions containing coated magnetite increased efficiency due to co-condensation of DNA. From 10(8) Escherichia coli cells, 7.1 microg of DNA was recovered using 100 microg of magnetite. E. coli DNA extracted with modified bacterial magnetite was suitable for restriction enzyme digestion and polymerase chain reaction (PCR). Ten replicate samples of E. coli cells were extracted using an automated magnetic robot and 9.5 microg of DNA was extracted using 100 microg of modified bacterial magnetite and possessed a 1.94 absorbance ratio (260:280 nm).
Yoza
Brandon
B
Department of Biotechnology, Tokyo University of Agriculture and Technology, 2-24-16 Koganei, Tokyo 184-8588, Japan.
Matsumoto
Mitsufumi
M
Matsunaga
Tadashi
T
D000588:Amines / Q000737:chemistry; D004269:DNA, Bacterial / Q000302:isolation & purification* / Q000378:metabolism; D004587:Electrophoresis, Agar Gel; D004926:Escherichia coli / Q000235:genetics*; D005029:Ethylenediamines / Q000737:chemistry; D052203:Ferrosoferric Oxide; D006863:Hydrogen-Ion Concentration; D007501:Iron / Q000737:chemistry* / Q000378:metabolism; D017646:Organosilicon Compounds / Q000737:chemistry*; D010087:Oxides / Q000737:chemistry* / Q000378:metabolism; D010316:Particle Size; D016133:Polymerase Chain Reaction; D011437:Propylamines; D012821:Silanes / Q000737:chemistry
D000588:Amines; D004269:DNA, Bacterial; D005029:Ethylenediamines; D017646:Organosilicon Compounds; D010087:Oxides; D011437:Propylamines; D012821:Silanes; C110457:3-(2-(2-aminoethyl)ethylamino)propyltrimethoxysilane; D007501:Iron; C477625:amino-propyl-triethoxysilane; D052203:Ferrosoferric Oxide