Patent ID: 6855496
Filing Date: 2005-02-15
Classification: A61K,B82Y,C07H,C07K,C12N,C12Q,C40B,F02B,G01N

Abstract:
1. A method of identifying a single stranded RNA ligand of a target molecule from a candidate mixture of single-stranded RNA molecules each comprising a region of randomized sequence, said method comprising: a) contacting said candidate mixture with said target molecule; b) partitioning the RNA molecules having an increased affinity to the target molecule relative to the candidate mixture from the remainder of the candidate mixture; c) adding a first tail sequence to said RNA molecules partitioned in step b) using terminal transferase; d) reverse transcribing said tailed RNA molecules to cDNA using a primer complementary to said first tail sequence; e) adding a second tail sequence to said cDNA molecule using terminal transferase, wherein said first and said second tail sequences have different sequences; f) amplifying said tailed cDNA molecules of e) using said primer of step d) and a primer comprising sequence complementary to said second tail sequence and further comprising an RNA polymerase promoter sequence, whereby double stranded DNA template is produced; g) transcribing said double stranded DNA template off) to yield single-stranded RNA molecules; and h) removing said tail sequences from said single-stranded RNA molecules of g) whereby a single-stranded RNA ligand of said target molecule is identified.