Patent ID: 9708603
Date: 2017-07-18
CPC Classifications: C12N,C12Q

Claim:
1. A method for amplifying cDNA from mRNA in a cell, comprising: (1) capturing mRNA derived from the cell by a first DNA probe comprising a first tag sequence and a poly T sequence and being immobilized to a solid carrier in a single reaction vessel, and synthesizing a first strand cDNA from the mRNA by a reverse transcription reaction; (2) removing a reaction reagent from the reaction vessel while keeping the first strand cDNA synthesized onto the solid carrier in the reaction vessel; (3) adding a polynucleotide sequence consisting of one type of nucleotides to 3′ terminal of the first strand cDNA on the solid carrier; (4) hybridizing a second DNA probe comprising a second tag sequence and a complementary sequence to the polynucleotide sequence with the cDNA to which the polynucleotide sequence is added, and synthesizing a second strand cDNA without a degradation and removal of the first DNA probe by a DNase reaction; and (5) performing a DNA amplification reaction using the second strand cDNA synthesized on the solid carrier as a template.