- 专利标题: Solid phase technique for selectively isolating nucleic acids
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申请号: US11129218申请日: 2005-05-13
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公开(公告)号: US20060003357A1公开(公告)日: 2006-01-05
- 发明人: Kevin McKernan , Paul McEwan , William Morris
- 申请人: Kevin McKernan , Paul McEwan , William Morris
- 申请人地址: US MA Cambridge 02142
- 专利权人: Whitehead Institute for Biomedical Research
- 当前专利权人: Whitehead Institute for Biomedical Research
- 当前专利权人地址: US MA Cambridge 02142
- 主分类号: C12Q1/68
- IPC分类号: C12Q1/68 ; C12N1/08
摘要:
A method of isolating target nucleic acid molecules from a solution comprising a mixture of different size nucleic acid molecules, in the presence or absence of other biomolecules, by selectively facilitating the adsorption of a particular species of nucleic acid molecule to the functional group-coated surface of magnetically responsive paramagnetic microparticles is disclosed. Separation is accomplished by manipulating the ionic strength and polyalkylene glycol concentration of the solution to selectively precipitate, and reversibly adsorb, the target species of nucleic acid molecule, characterized by a particular molecular size, to paramagnetic microparticles, the surfaces of which act as a bioaffinity adsorbent for the nucleic acids. The target nucleic acid is isolated from the starting mixture based on molecular size and through the removal of magnetic beads to which the target nucleic acid molecules have been adsorbed. The disclosed method provides a simple, robust and readily automatable means of nucleic acid isolation and purification which produces high quality nucleic acid molecules suitable for: capillary electrophoresis, nucleotide sequencing, reverse transcription cloning the transfection, transduction or microinjection of mammalian cells, gene therapy protocols, the in vitro synthesis of RNA probes, cDNA library construction and PCR amplification.
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