发明公开
- 专利标题: ENDOTOXIN REDUCTION IN NUCLEIC ACID PURIFICATION
- 专利标题(中): 内毒素在DNA清洗
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申请号: EP99918650.5申请日: 1999-04-22
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公开(公告)号: EP1071695A1公开(公告)日: 2001-01-31
- 发明人: SMITH, Craig, E. , CRESWELL, Donald, A. , BITNER, Rex, M. , WHITE, Douglas, H. , BUTLER, Braeden, L. , LESLEY, Scott, A.
- 申请人: PROMEGA CORPORATION
- 申请人地址: 2800 Woods Hollow Road Madison, WI 53711-5399 US
- 专利权人: PROMEGA CORPORATION
- 当前专利权人: PROMEGA CORPORATION
- 当前专利权人地址: 2800 Woods Hollow Road Madison, WI 53711-5399 US
- 代理机构: Watson, Robert James
- 优先权: US64449 19980422
- 国际公布: WO9954340 19991028
- 主分类号: C07H21/00
- IPC分类号: C07H21/00 ; C07H21/02 ; C07H21/04 ; C12Q1/68
摘要:
The present invention presents a novel method for removing endotoxins from nucleic acids, such as DNA, RNA, or hybrids thereof, contaminated therewith. Nucleic acid solutions which can be treated using the method of this invention include, but are not limited to, lysates of gram-negative bacteria and nucleic acid solutions contaminated with endotoxins from external sources. The present method removes endotoxins from such solutions using silica-based materials, such as silica gel particles, magnetic silica particles, or diatomaceous earth. In a preferred aspect of the method of this invention, magnetic silica particles are used to isolate plasmid DNA from a lysate of gram-negative bacteria transformed with the plasmid DNA. Application of the disclosed method produces nucleic acids which are sufficiently free of endotoxin contamination to be useful for a variety of different practical applications. Figure 1 shows a plot of units of endotoxin bound vs the number of milligrams of particles added to lysates of each of two differerent strains of bacteria, JM109 (◊) or DH5α(♦).
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