Patent ID: 6955876

Claim:
A method for identification and characterization of gene expression in one or more samples, comprised of: (a) providing one or more samples comprising one or more mRNA molecules; (b) providing an identimer comprising an oligo-dT primer of sequence, from 5′ to 3′ end, of Tn-VNx, where n is an integer 8 or greater but not more than 50 representing the number of T's, V equals a nucleotide A, C, or G but not T, each N equals a nucleotide A, C, G, or T, and x is an integer 3 or greater but not more than 10 representing the number of N nucleotides, said identimer also comprising a detectable marker at its 5′ end; (c) contacting said mRNA with said identimer such that the polyT portion of the identimer hybridizes to the polyA tail of the mRNA and the VNx portion of the identimer hybridizes with portions of the mRNA immediately upstream of the polyA tail; (d) reverse transcribing the mRNA to produce a first strand cDNA that includes the identimer; (e) synthesizing a second DNA strand complementary to the first strand cDNA to form a duplex; (f) cleaving the duplex with at least one sequence-specific cleaving agent to provide one or more duplex cleavage fragments; (g) ligating an adaptamer comprising an RNA polymerase promoter site to one or more of said cleavage fragments; and (h) amplifying the one or more ligated cleavage fragments using the identimer to produce one or more amplified fragments comprising sequences complementary to a 3′ end of the mRNA.