Template-dependent nucleic acid polymerization using oligonucleotide triphosphates building blocks

• US 7,060,440 B1
• Filed: 08/29/2000
• Issued: 06/13/2006
• Est. Priority Date: 09/01/1999
• Status: Expired due to Term

• Current Assignee: Yeda Research and Development Co. Ltd. (Weizmann Institute of Science)
• Original Assignee: Nuace Technologies Ltd., Yeda Research and Development Co. Ltd. (Weizmann Institute of Science)
• Inventors: Kless, Hadar
• Primary Examiner(s): Horlick, Kenneth R.
• Assistant Examiner(s): WILDER, CYNTHIA B
• Application Number:
  US10/069,236
• Time in Patent Office: 2,114 Days
• Field of Search: 435/6, 435/91.1, 435/91.2, 435/91.21, 536/23.1, 536/24.3, 536/24.32, 536/24.33
• US Class Current: 435/6.1
• CPC Class Codes: C12N 9/1252 DNA-directed DNA polymerase (2.7.7.7), i.e. DNA replicase
• CPC Class Codes: C12P 19/34 Polynucleotides, e.g. nucleic acids, oligoribonucleotides
• CPC Class Codes: C12Q 1/6844 Nucleic acid amplification reactions
• CPC Class Codes: C12Q 1/686 Polymerase chain reaction [PCR]
• CPC Class Codes: C12Q 1/6869 Methods for sequencing
• CPC Class Codes: C12Q 2521/325 Single stranded exonuclease
• CPC Class Codes: C12Q 2525/204 specific length of the oligonucleotides
• CPC Class Codes: C12Q 2533/101 Primer extension

Abstract

A novel use of a template-dependent polymerase. The novel use is effected by employing the template-dependent polymerase for incorporating at least one oligonucleotide triphosphate onto a nascent oligonucleotide-3″-OH in a template pendent manner.

Claims

1. A method of extending a nascent oligonucleotide-3′

-OH in a template-dependent manner, the method comprising the step of contacting the nascent oligonucleotide-3′

-OH with a nucleic acid template, a template-dependent polymerase and 4^N oligonucleotide triphosphates each including N monomers, wherein N is an integer equal to 2 or 3 under conditions in which said nascent oligonucleotide-3′

-OH hybridizes with said nucleic acid template and said template-dependent polymerase is active in incorporating said oligonucleotide triphosphates onto a growing 3′

-OH group of the nascent oligonucleotide-3′

-OH, thereby extending the nascent oligonucleotide-3′

-OH in a template-dependent manner.