Patent ID: 9416406
Filing Date: 2016-08-16
CPC Classification: C12N,C12Q

Claim Text:
1. A method for detecting an RNA molecule, comprising: (a) forming a single ligation reaction mixture containing (i) at least one RNA molecule including at least a first and a second RNA molecule, (ii) a plurality of a first double-stranded nucleic acid adaptor, (iii) a plurality of a second double-stranded nucleic acid adaptor, and (iv) an RNA ligase, wherein a first strand of the first double-stranded nucleic acid adaptors of the plurality include at least two ribonucleosides at the 3′ end, and a second strand of the first double-stranded nucleic acid adaptors of the plurality include a first single-stranded portion at the 5′ end, and wherein a first strand of the second double-stranded nucleic acid adaptors of the plurality include a terminal 5′ phosphate group, and a second strand of the second double-stranded nucleic acid adaptors of the plurality include a second single-stranded portion at the 3′ end, and wherein the first and the second single-stranded portions contain a nucleotide sequence that hybridizes to a portion of the at least first RNA molecule from the mixture of different RNA molecules, and wherein the first double-stranded adaptors of the plurality, or the second double-stranded adaptors of the plurality, or the first and second double-stranded adaptors comprise a unique bar code sequence; and (b) producing a plurality of different RNA ligation product by ligating the first double-stranded adaptor to a first end of the at least one RNA molecule and ligating the second double-stranded adaptor to a second end of the at least one RNA molecule.