C - Chemistry – Metallurgy – 07 – H
Patent
C - Chemistry, Metallurgy
07
H
C07H 21/00 (2006.01) C07H 21/02 (2006.01) C07H 21/04 (2006.01) C12N 15/10 (2006.01) C12Q 1/68 (2006.01)
Patent
CA 2329067
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 DH5.alpha.(♦).
L'invention concerne un nouveau procédé servant à supprimer des endotoxines d'acides nucléiques, tels qu'ADN, ARN ou leurs hybrides, contaminés par ces endotoxines. Des solutions d'acides nucléiques qu'on peut traiter au moyen de ce procédé comprennent, sans y être toutefois limitées, des lysats de bactéries à Gram négatif et des solutions d'acides nucléiques contaminées par des endotoxines provenant de sources extérieures. Ce procédé permet de supprimer les endotoxines de ces solutions au moyen de matériaux à base de silice, tels que des particules de gel de silice, des particules de silice magnétique ou de la terre de diatomées. Dans un aspect préféré de ce procédé, on utilise des particules de silice magnétique afin d'isoler l'ADN de plasmides depuis un lysat de bactéries à Gram négatif transformées par cet ADN plasmidique. La mise en application de ce procédé permet d'obtenir des acides nucléiques suffisamment exempts de contamination par les endotoxines pour servir à une variété de différents domaines d'utilisation pratique.
Bitner Rex M.
Butler Braeden L.
Creswell Donald A.
Lesley Scott A.
Smith Craig E.
Promega Corporation
Shapiro Cohen
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