公开(公告)号：US20190276883A1

公开(公告)日：2019-09-12

申请号：US16302485

申请日：2017-05-17

Applicant: Japan Science and Technology Agency

Inventor： Masayuki Su'etsugu ,  Hiroko Tsujimoto ,  Takeshi Shinohara

IPC: C12Q1/6855

Abstract: Provided is a method capable of simply and exponentially amplifying circular DNA, and particularly, long-chain circular DNA, in a cell-free system. Specifically, provided herein is a method for amplifying circular DNA which comprises mixing circular DNA having a replication origin sequence (origin of chromosome (oriC)) with a reaction solution comprising: a first enzyme group that catalyzes replication of circular DNA; a second enzyme group that catalyzes an Okazaki fragment maturation and synthesizes two sister circular DNAs constituting a catenane; a third enzyme group that catalyzes a separation of two sister circular DNAs; and also, a buffer, NTP, dNTP, a magnesium ion source, and an alkali metal ion source, to form a reaction mixture, which is then reacted.