Method of polyadenylation and CDNA synthesis in a single reaction

US9217173B2 · kind B2 · utility

Assignee

• QIAGEN GmbH · DE

Inventors

• Holger Engel · Hilden, DE
• Subrahmanyam Yerramilli · Clarksville, US
• Martin Kreutz · Germantown, US
• Dirk Loeffert · Haan, DE
• Christian Korfhage · Hilden, DE

Key dates

• Filing date: Aug 13, 2007
• Grant date: Dec 22, 2015
• Expiry date: Apr 2, 2032

Classification

• Technology area (CPC C): Chemistry; Metallurgy
• CPC primary: C12Q1/6848
• WIPO field: Biotechnology
• WIPO sector: Chemistry

Abstract

This invention relates to a process for synthesis of a cDNA in a sample, in an enzymatic reaction, wherein the process comprises the steps: simultaneous preparation of a first enzyme with polyadenylation activity, a second enzyme with reverse transcriptase activity, a buffer, at least one ribonucleotide, at least one deoxyribonucleotide, an anchor oligonucleotide; addition of a sample that comprises a ribonucleic acid; and incubation of the agents of the previous steps in one or more temperature steps, which are selected such that the first enzyme and the second enzyme show activity. The invention further relates to a reaction mixture that comprises a first enzyme with polyadenylation activity, a second enzyme with reverse transcriptase activity, optionally a buffer, optionally at least one ribonucleotide, optionally at least one deoxyribonucleotide, and optionally an anchor oligonucleotide. Moreover, the invention relates to a kit that comprises a corresponding reaction mixture.