公开(公告)号：US20220356462A1

公开(公告)日：2022-11-10

申请号：US17625918

申请日：2020-07-10

Applicant: TOKYO UNIVERSITY OF SCIENCE FOUNDATION

Inventor： Kouji MATSUSHIMA ,  Satoshi UEHA ,  Shigeyuki SHICHINO ,  Satoru ITO ,  Hiroyasu AOKI

IPC: C12N15/10 ,  C12Q1/6874

Abstract: A nucleic acid amplification method using a solid-phase carrier according to the present invention comprises: capturing a target nucleic acid comprising mRNA on a solid-phase carrier; carrying out complementary-strand synthesis on the solid phase; carrying out exonuclease treatment to degrade and remove unreacted target- capturing nucleic acid on the solid phase; and then carrying out mRNA degradation and homopolymer addition by TdT reaction in the presence of a chain-terminating nucleotide triphosphate. According to the method of the present invention, cDNA can be stably and highly efficiently amplified even from a small amount of sample even in cases where the ratio of the amount of enzyme to the DNA substrate on the solid phase is excessive, where the reaction time is excessive, and/or where reagents show lot-to-lot variation. Further, the amplification method of the present invention can broaden the range of applications of techniques in which analysis using a specific-binding molecule labeled with an oligonucleic acid such as a DNA-labeled antibody and analysis of transcripts are carried out simultaneously.