Patent ID: 9222129
Filing Date: 2015-12-29
Classification: C12Q

Abstract:
1. A method for preparing a group of nucleic acids comprising amplification products retaining a relative relation of each gene expression level in a biological sample, comprising: (a) preparing a primary cDNA chain by reverse transcription using mRNA isolated from a biological sample as a template and using a first primer comprising a poly(T) sequence and an additional nucleotide sequence X thereto; (b) inactivating the remaining first primer after the reaction in (a); (c) subjecting the primary cDNA chain obtained in (a) to a poly(A) tailing reaction followed by preparing a secondary double-stranded DNA chain using the poly(A) tailing reaction product as a template and using a second primer comprising poly(T) sequence and an additional nucleotide sequence Y thereto, wherein the nucleotide sequences X and Y in the first and second primers, respectively, have different sequences from each other; (d) adding the first primer, and performing PCR amplification between the first primer and the second primer; and (e) performing PCR amplification using primer set (1) or (2) below, wherein the nucleotide sequences X and Y do not contain promoter sequences that are recognized by RNA polymerase.