Patent ID: 6864050

Claim:
A method for the amplification of a population of nucleic acids comprising a population of poly(A)+ RNA, wherein said method preserves the relative abundance of individual nucleic acid species, said method comprising: a first step of mixing said population of nucleic acids with a primer comprising oligo-dT in a single reaction vessel under conditions that allow hybridization of said primer with said population of poly(A)+ RNA; a second step of synthesizing a single-stranded DNA population from said population of poly(A)+ RNA wherein a reverse transcriptase, dNTPs and a first buffer are added to the single reaction vessel to synthesize said single-stranded DNA population; a third step of synthesizing a population of double-stranded DNA from said single-stranded DNA population wherein a second buffer, different from said first buffer, and a four enzyme-mix comprising a DNA polymerase are added to said single reaction vessel to synthesize said double-stranded cDNA and wherein the reaction is incubated at about 16° C. for at least about 2 hours followed by heating to about 75° C. for at least 1 minute; and a fourth step of synthesizing multiple copies of RNA from said double-stranded DNA population, wherein an RNA polymerase and a third buffer, different from said first and second buffers, are added to said single reaction vessel to synthesize said multiple copies of RNA.