Search International and National Patent Collections

1. (WO2016092062) METHOD FOR IMPROVING THE EFFICIENCY OF CDNA GENERATION

Pub. No.:    WO/2016/092062    International Application No.:    PCT/EP2015/079375
Publication Date: Jun 16, 2016 International Filing Date: Dec 11, 2015
IPC: C12Q 1/68
C12N 15/10
Applicants: QIAGEN GMBH
Inventors: FANG, Nan
LOEFFERT, Dirk
Title: METHOD FOR IMPROVING THE EFFICIENCY OF CDNA GENERATION
Abstract:
The present invention relates to a method for synthesizing, in an enzymatic reaction, a cDNA from a degraded and/or modified ribonucleic acid template present in a sample, as well as to the use of this method, wherein the method comprises the following steps: a. providing a reaction mixture comprising a first enzyme with polyadenylation activity, a second enzyme with reverse transcriptase activity, a buffer, adenosine-5'-triphosphates (rATP), deoxyribonucleotides (dNTP), oligo dT-primers, and random primers; b. adding a sample comprising a degraded and/or modified ribonucleic acid template to the reaction mixture from step a; and c. incubating the reaction mixture from step a. with the sample of step b. in one or more temperature steps, which are selected so that the first enzyme and the second enzyme display activity.