期刊
JOURNAL OF BIOTECHNOLOGY
卷 94, 期 3, 页码 217-224出版社
ELSEVIER SCIENCE BV
DOI: 10.1016/S0168-1656(01)00427-8
关键词
DNA extraction; bacterial magnetite; 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 mug of DNA was recovered using 100 kg 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 mug of DNA was extracted using 100 mug of modified bacterial magnetite and possessed a 1.94 absorbance ratio (260:280 nm). (C) 2002 Elsevier Science B.V. All rights reserved.
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