Method for producing circular DNA formed from single-molecule DNA

摘要

There is provided a method for producing a circular DNA which consists of a circular DNA formed from a single-molecule DNA and which does not comprise circular DNA formed from multiple-molecule DNA. According to the method of the present invention, a circular DNA molecule formed only from a single-molecule DNA can be reliably produced.

主权项

1. An adapter for producing a circular DNA, the adapter consisting of two DNA strands complementary to each other having the following structure y1-Y-y2-X: wherein X represents a double-stranded DNA being a restriction enzyme recognition site comprising a palindromic sequence; y1 and y2 represent double-stranded DNAs respectively comprising identical nick-generating enzyme recognition sites being reversely oriented to each other; Y represents a double-stranded DNA sequence having a unique sequence for each DNA molecule to be circularized, n represents an integer of 4 to 15, N1 to Nn, which may be the same or different, each represent a deoxyribonucleotide selected from the group consisting of dAMP, dCMP, dGMP and dTMP, and N′1 to N′n represent the following deoxyribonucleotides respectively corresponding to the N1 to Nn,TABLE 1NkN′kdAMPdTMPdCMPdGMPdGMPdCMPdTMPdAMP wherein k represents an integer from 1 to n.