| 摘要 |
The problem of the present invention is to provide a method for preparing a specimen for the comprehensive and accurate analysis of gene expression in one or few cells using a large-scale DNA sequencer. The present invention relates to a method for amplifying cDNA from mRNA in a cell, the method comprising (1) a step for, in one reactor, capturing the cell-derived mRNA using a first DNA probe formed from a first tag sequence and a poly T sequence and immobilized on a solid carrier and synthesizing a first strand cDNA from the mRNA by reverse transcription, (2) a step for removing the reaction reagent from the reactor vessel in which the first strand cDNA synthesized on the solid carrier remains, (3) a step for adding a polynucleotide sequence consisting of one nucleotide to the 3' terminal of the first strand cDNA on the solid carrier, (4) a step for hybridizing a second DNA probe containing a second tag sequence and a sequence complementary to the polynucleotide sequence with cDNA to which the polynucleotide sequence has been added and synthesizing a second strand cDNA, and (5) a step for performing a DNA amplification using the second strand cDNA synthesized on the solid carrier as the template. |
