Patent ID: 7361465

Claim:
A method for amplifying a targeted RNA in a sample comprising: (a) altering the secondary structure of the targeted RNA by denaturing the targeted RNA or contacting the targeted RNA with a single-stranded binding protein; (b) incubating the targeted RNA in the presence of poly(A) polymerase I of E. coli and a nucleotide under conditions that allow tailing of the targeted RNA wherein said conditions include a buffer lacking Mn 2+ and, for RNA amounts in the range of 1-100 ng, the nucleotide concentration is 5-350 μM; (c) hybridizing the targeted RNA with a promoter-oligo-dN primer complementary to the tail of the targeted RNA; (d) extending the promoter-oligo-dN primer using a reverse transcriptase to form a first strand complementary to the targeted RNA; (e) synthesizing a second strand complementary to the first strand; and (f) transcribing copies of RNA initiated from the promoter-oligo-dN primer using an RNA polymerase, wherein the RNA is complementary to the second strand.