Patent ID: 7432084
Filing Date: 2008-10-07
Classification: C12N,C12Q

Abstract:
1. A method of synthesizing a preparation of nucleic acid molecules, the method comprising the steps of: (a) utilizing an RNA template to enzymatically synthesize a population of first DNA molecules that are complementary to at least 50 contiguous bases of said RNA template, wherein the synthesis of the population of first DNA molecules is primed using a first primer mixture comprising a multiplicity of first primers, wherein each of the first primers comprises a random sequence portion, and a defined sequence portion; (b) utilizing the population of first DNA molecules as a template to enzymatically synthesize a population of second DNA molecules, thereby forming a population of double-stranded DNA molecules wherein the population of first DNA molecules is hybridized to the population of second DNA molecules; (c) utilizing the population of first or second DNA molecules of the double-stranded DNA molecules as a template to enzymatically synthesize a population of first RNA molecules that are complementary to either the first DNA molecules or to the second DNA molecules; (d) utilizing the population of first RNA molecules as a template to enzymatically synthesize a population of third DNA molecules that are complementary to the first RNA molecules; and (e) labeling the population of third DNA molecules with at least one dye molecule.