Methods for making nucleic acids

US6582936B1 · kind B1 · utility

Assignee

• The Regents of the University of California · US

Inventors

• Tito Serafini · San Francisco, US
• Percy Luu · Oakland, US
• John Ngai · Boxborough, US
• David M. Lin · Ithaca, US

Key dates

• Filing date: May 8, 2000
• Grant date: Jun 24, 2003
• Expiry date: May 8, 2020

Classification

• Technology area (CPC C): Chemistry; Metallurgy
• CPC primary: C12Q1/6809
• WIPO field: Biotechnology
• WIPO sector: Chemistry

Abstract

Nucleic acids are made by converting a primed single-stranded DNA to a double-stranded DNA by a method comprising the step contacting the single-stranded DNA with a DNA polymerase having 5′ exonuclease activity under conditions whereby the DNA polymerase converts the-single stranded DNA to the double-stranded DNA, wherein the single-stranded DNA is primed with oligonucleotide primer comprising a sequence complementary to the 3′ end of the single-stranded DNA, and at least one of the 5′ end of the primer and the single-stranded DNA comprises an RNA polymerase promoter joined to an upstream (5′) flanking moiety which protects the promoter from the 5′ exonuclease activity of the DNA polymerase.