Invention Grant
- Patent Title: Amplification and cloning of single DNA molecules using rolling circle amplification
- Patent Title (中): 使用滚环扩增扩增和克隆单个DNA分子
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Application No.: US11919515Application Date: 2006-05-01
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Publication No.: US08497069B2Publication Date: 2013-07-30
- Inventor: Clyde A. Hutchison, III , Hamilton O. Smith
- Applicant: Clyde A. Hutchison, III , Hamilton O. Smith
- Applicant Address: US CA San Diego
- Assignee: Synthetic Genomics, Inc.
- Current Assignee: Synthetic Genomics, Inc.
- Current Assignee Address: US CA San Diego
- Agency: DLA Piper LLP (US)
- International Application: PCT/US2006/016349 WO 20060501
- International Announcement: WO2006/119066 WO 20061109
- Main IPC: C12Q1/68
- IPC: C12Q1/68 ; C12P19/34

Abstract:
The present invention relates, e.g., to a method for amplifying a small number of copies (e.g. a single copy) of a single-stranded circular DNA molecule (e.g. having a size of about 5-6 kb) by an isothermal rolling circle mechanism, using random or partially random primers and a F29-type DNA polymerase. The method, which can also be used for amplifying DNAs by non-rolling types of multiple displacement amplification, comprises incubating the reaction components in a small volume, e.g. about 10 μl or less, such as about 0.6 μl or less. The degree of amplification can be about 109 fold, or higher. A method for cell-free cloning of DNA, using the rolling circle amplification method of the invention, is described.
Public/Granted literature
- US20090176280A1 Amplification and cloning of single dna molecules using rolling circle amplification Public/Granted day:2009-07-09
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