Method for derivitization of polynucleotides

• US 4,739,044 A
• Filed: 06/13/1985
• Issued: 04/19/1988
• Est. Priority Date: 06/13/1985
• Status: Expired due to Term

• Current Assignee: Amgen Inc.
• Original Assignee: Amgen Inc.
• Inventors: Stabinsky, Yitzhak
• Primary Examiner(s): Brown, J. R.
• Assistant Examiner(s): Rollins, John W.
• Application Number:
  US06/744,798
• Time in Patent Office: 1,041 Days
• Field of Search: 536/27, 536/28, 536/29
• US Class Current: 536/25.3
• CPC Class Codes: C07H 21/00 Compounds containing two or more mononucleotide units having separate phosphate or polyphosphate groups linked by saccharide radicals of nucleoside groups, e.g. nucleic acids

Abstract

A method for the preparation of a 3'"'"' end functionalized polynucleotide is disclosed. An amine-functionalized solid phase support is treated sequentially with an anhydride, then with an ω-hydroxylamine. A polynucleotide is chemically synthesized on the treated support and is subsequently cleaved therefrom by hydrolysis of the amide bonds. A polynucleotide having a 3'"'"' free primary amine is recovered for use in hybridization assays and other uses.

Claims

1. A method for preparing a functionalized polynucleotide comprising the steps of:

forming an amide bond between a support-bound carboxyl group and an amino group of a hydroxylamine linker molecule;

initiating synthesis of a polynucleotide on the hydroxyl group of the hydroxylamine linker molecule;

hydrolyzing the amide bound between the amine and the support-bound carboxyl group; and

recovering the polynucleotide connected to the amine group by way of the linker molecule.

2. The method according to claim 1 wherein said polynucleotide sequence is a deoxyribonucleic acid sequence.

3. The method according to claim 1 wherein said polynucleotide sequence is a ribonucleic acid sequence.

4. The method according to claim 1 wherein said hydroxylamine contains from 1 to 10 carbon atoms.

5. The method according to claim 4 wherein said hydroxylamine is selected from the group comprising methanolamine, ethanolamine, propanolamine, butanolamine, pentanolamine, hexanolamine, heptanolamine, octanolamine, nonanolamine and decanolamine.