Method for the uncoupled, direct, exponential amplification and sequence of DNA molecules with the addition of a second thermostable DNA polymerase and its application

US6225092A · kind A · utility

Assignee

• Roche Diagnostics GMBH · DE

Inventors

• Christian Kilger · Hersbruck, DE
• Svante Paabo · Hersbruck, DE

Key dates

• Filing date: Dec 16, 1997
• Grant date: May 1, 2001
• Expiry date: Dec 16, 2017

Classification

• Technology area (CPC Y): Emerging Cross-Sectional Technologies
• CPC primary: Y10S435/81
• WIPO field: Biotechnology
• WIPO sector: Chemistry

Abstract

Method for sequencing a nucleic acid molecule in a thermocycling reaction which initially comprises a nucleic acid molecule, a first primer, a second primer, a reaction buffer, a first thermostable DNA polymerase, (optionally) a thermostable pyrophosphatase, deoxynucleotides or derivatives thereof and a dideoxynucleotide or a derivative thereof and which is characterized in that the thermocycling reaction additionally contains a second thermostable DNA polymerase which, in comparison to the said first thermostable DNA polymerase, has a reduced ability to incorporate dideoxynucleotides as well as the use of the said method.