Polycationic buffers and method for gel electrophoresis of nucleic acids

• US 4,936,963 A
• Filed: 05/23/1989
• Issued: 06/26/1990
• Est. Priority Date: 05/27/1987
• Status: Expired due to Fees

• Current Assignee: Abbott Laboratories
• Original Assignee: Abbott Laboratories
• Inventors: Hayden, Mark A., Mandecki, Wlodek
• Primary Examiner(s): Niebling, John F.
• Assistant Examiner(s): STARSIAK, JOHN S
• Application Number:
  US07/356,590
• Time in Patent Office: 399 Days
• Field of Search: 204/182.8, 204/299 R, 204/180.1
• US Class Current: 204/468
• CPC Class Codes: G01N 27/44747 Composition of gel or of carrier mixture

Abstract

The present invention provides for polycationic buffers for the gel electrophoresis of and methods for obtaining improved separation and resolution of nucleic acids. In particular, the invention provides for a buffer consisting essentially of histidine for the gel electrophoresis of oligonucleotides ranging from a few to about 70 nucleotides in length. Polycationic buffers may be used for both analytical and preparative purposes and result in a nearly two-fold improvement in both separation and resolution of the nucleic acid bands. Furthermore, the use of polycationic buffers reduces the electrophoresis time by a factor of about three due to the low conductivity of the buffers.

Claims

1. A method for the gel electrophoresis of nucleic acids comprising the steps of:

introducing a polycationic buffer to a gel;

applying a sample of a nucleic acid on said gel; and

applying an electromotive potential difference across said gel until resolution of said sample of a nucleic acid into its component parts.

2. The method as recited in claim 1 wherein the polycationic buffer is selected from the group consisting of polyamines, polyamino acids and aliphatic polyamines.

3. The method as recited in claim 1 wherein said step of introducing a polycationic buffer comprises the step of applying an aqueous solution of histidine at a concentration within a range of from 10 mM to 100 mM.

4. The method as recited in claim 1 wherein said step of applying a sample of denatured nucleic acid on a gel comprises the step of preparing a gel which is a polyacrylamide gel having a concentration of from 8% to 20% cross-linking.