Reaction mixtures for forming cDNA from an RNA template

US9611506B2 · 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: Nov 12, 2015
• Grant date: Apr 4, 2017
• Expiry date: Nov 12, 2035

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, whereby 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.