Patent ID: 7354742

Claim:
A method for amplifying at least one mRNA in a sample containing a plurality of different mRNAs comprising: a) synthesizing first strand cDNA by contacting under conditions conducive to reverse transcriptase activity at least one mRNA in said sample with (i) reverse transcriptase, and (ii) a first primer that is sufficiently complementary to a sequence in the mRNA so as to prime synthesis in a direction toward the 5â€² end of the mRNA; b) synthesizing a second strand of cDNA by contacting under conditions conducive to a thermostable DNA polymerase activity, said conditions comprising an incubation temperature of from 55Â° C. to 70Â° C., the first strand cDNA with (i) a thermostable DNA polymerase, which is Bst DNA polymerase large fragment, and (ii) a thermostable RNase H, thereby forming double stranded cDNA; and c) transcribing the double stranded cDNA into cRNA by contacting the double stranded cDNA with an RNA polymerase under conditions conducive to RNA polymerase activity, such that cRNA is produced.