Patent ID: 8192941
Filing Date: 2012-06-05
Classification: C12N,C12Q

Abstract:
1. A method for detecting a RNA molecule in a sample, comprising: combining the sample with at least one first adaptor, at least one second adaptor, and a polypeptide comprising double-strand specific RNA ligase activity to form a ligation reaction composition in which the at least one first adaptor and the at least one second adaptor are ligated to the RNA molecule of the sample to form a ligated product in the same ligation reaction composition, combining the ligated product with reverse transcribing the ligated product to form a reverse transcribed product, digesting at least some of the ribonucleosides from the reverse transcribed product with ribonuclease H to form an amplification template, combining the amplification template with at least one forward primer, at least one reverse primer, and a DNA-directed DNA polymerase when the ligated product is combined as in i), to form an amplification reaction composition, cycling the amplification reaction composition to form at least one amplified product, and determining the sequence of at least part of the amplified product, thereby detecting the RNA molecule.