Method for producing circular DNA formed from single-molecule DNA

• US 9,434,941 B2
• Filed: 01/12/2015
• Issued: 09/06/2016
• Est. Priority Date: 09/02/2010
• Status: Active Grant

• Current Assignee: Kurume University
• Original Assignee: Kurume University
• Inventors: Mizuno, Shinichi, Nagafuji, Koji, Okamura, Takashi
• Primary Examiner(s): Thomas, David
• Application Number:
  US14/594,458
• Publication Number:
  US 20150197745A1
• Time in Patent Office: 603 Days
• Field of Search: None
• US Class Current: 1/1
• CPC Class Codes: C07H 21/04 with deoxyribosyl as saccharide radical
• CPC Class Codes: C12N 15/10 Processes for the isolation, preparation or purification of DNA or RNA chemical preparation of DNA or RNA C07H21/00; preparation of non-structural polynucleotides from microorganisms or with enzymes C12P19/34
• CPC Class Codes: C12N 15/1096 cDNA Synthesis; Subtracted cDNA library construction, e.g. RT, RT-PCR
• CPC Class Codes: C12N 15/64 General methods for preparing the vector, for introducing it into the cell or for selecting the vector-containing host
• CPC Class Codes: C12Q 1/6806 Preparing nucleic acids for analysis, e.g. for polymerase chain reaction [PCR] assay C12Q1/6804 takes precedence
• CPC Class Codes: C12Q 2525/307 Circular oligonucleotides

Abstract

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.

Claims

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:

2. The adapter according to claim 1, wherein the nick-generating enzyme is Nb.BtsI or Nt.BsqQI.

3. A kit for producing circular DNA, comprising an adapter consisting of a double-stranded DNA comprising a restriction enzyme recognition site identical to the restriction enzyme recognition site represented by X defined in claim 1, and the adapter according to claim 1.