Enhancement of the transfection efficiency of nucleic acids by using isopropanol in aqueous solutions

• US 5,792,651 A
• Filed: 10/11/1996
• Issued: 08/11/1998
• Est. Priority Date: 02/07/1994
• Status: Expired due to Term

• Current Assignee: Qiagen GmbH (Qiagen NV)
• Original Assignee: Qiagen GmbH (Qiagen NV)
• Inventors: Schorr, Joachim, Colpan, Metin
• Primary Examiner(s): KETTER, JAMES S
• Application Number:
  US08/687,530
• Time in Patent Office: 669 Days
• Field of Search: 435/220, 536/25.41
• US Class Current: 435/270
• CPC Class Codes: C12N 15/101 by chromatography, e.g. electrophoresis, ion-exchange, reverse phase
• CPC Class Codes: C12N 15/87 Introduction of foreign genetic material using processes not otherwise provided for, e.g. co-transformation

Abstract

Use of isopropanol in aqueous solutions for chromatographic isolation of nucleic acids for enhancing the transfection efficiency of the isolated nucleic acids in prokaryotic and eukaryotic cells.

Claims

1. In a method of chromatographic isolation of nucleic acids, whereby nucleic acids are eluted using an aqueous solution, the improvement comprising eluting the nucleic acids using isopropanol in aqueous solution, which effects isolated nucleic acids having enhanced transfection efficiency in procaryotic or eucaryotic cells.

2. The method of claim 1 wherein said isopropanol is present in said aqueous solutions in an amount of from 1 to 50% by volume.

3. The method of claim 1 wherein said isopropanol is present in said aqueous solutions in an amount of from 5 to 25% by volume.

4. The method of claim 1 wherein said isopropanol is present in said aqueous solutions in an amount of from 10 to 15% by volume.

5. The method of claim 1 wherein said aqueous solution contains salts of monovalent or divalent cations.

6. The method of claim 5, wherein said aqueous solution contains sodium chloride, calcium chloride, potassium chloride, or combination thereof.

7. The method of claim 1 wherein said aqueous solution is a buffered solution.

8. The method of claim 1 wherein said nucleic acid DNA is plasmid DNA.

9. The method of claim 1 for the preparation of nucleic acids for gene therapy.

10. An aqueous solution containing 0.5 to 1.5M sodium chloride or potassium chloride, 10 to 100 mM MOPS or TRIS/HCl, and from 5 to 60% by volume of isopropanol at a pH value of 6 to 8.

11. An aqueous solution containing 1.0 to 2.0M sodium chloride or potassium chloride, 10 to 100 mM TRIS/HCl, and from 5 to 60% by volume of isopropanol at a pH value of 8 to 9.