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Method of isolating and purifying nucleic acids from biological samples

  • US 4,935,342 A
  • Filed: 12/01/1986
  • Issued: 06/19/1990
  • Est. Priority Date: 12/01/1986
  • Status: Expired due to Term
First Claim
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1. The method of isolating and purifying cellular or viral nucleic acids from an aqueous solution of a biological sample containing nuclein acids together with water-soluble contamination components of lysed cells or virus selected from the group consisting of proteins, pigments, carboxylated mucopolysaccharides, sulfated mucopolysaccharides, and mixtures thereof, comprising:

  • (a) selecting an anion exchange column material which effectively binds the target nucleic acids at a lower salt molarity than the molarity at which the target nucleic acids elute therefrom;

    (b) applying a sample containing nucleic acids together with water-soluble contaminating components of lysed cells or virus selected from the group consisting of proteins, pigments, carboxylated mucopolysaccharides, sulfated mucopolysaccharides, and mixtures thereof, to a column packed with the selected anion exchanger material, and said nucleic acids becoming bound to said column material;

    (c) washing said column with an aqueous solution of a salt at chloride molarity at which the nucleic acids remain bound to said column material, said wash molarity being close to the lowest molarity at which the nucleic acids begin to elute, said washing being of sufficient volume to wash the non-binding components through said column, including carboxylated mucopolysaccharides;

    (d) eluting the bound nucleic acids by passing through said column an aqueous solution of a salt having an eluting molarity corresponding to the lowest molarity at which the target nucleic acids will completely elute, said eluant being passed in sufficient volume to remove said nucleic acids from the column while leaving contaminating components including sulfated mucopolysaccharides in the column; and

    (e) recovering the eluted nucleic acids essentially free of mucopolysaccharides and other contaminating components.

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