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Title: Amplification and cloning of single DNA molecules using rolling circle amplification

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.

Inventors:
;
Issue Date:
Research Org.:
Synthetic Genomics, Inc., San Diego, CA (United States)
Sponsoring Org.:
USDOE
OSTI Identifier:
1531861
Patent Number(s):
8497069
Application Number:
11/919,515
Assignee:
Synthetic Genomics, Inc. (San Diego, CA)
Patent Classifications (CPCs):
C - CHEMISTRY C12 - BIOCHEMISTRY C12N - MICROORGANISMS OR ENZYMES
C - CHEMISTRY C12 - BIOCHEMISTRY C12Q - MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS
DOE Contract Number:  
FG02-02ER63453
Resource Type:
Patent
Resource Relation:
Patent File Date: 2006-05-01
Country of Publication:
United States
Language:
English

Citation Formats

Hutchison, III, Clyde A., and Smith, Hamilton O.. Amplification and cloning of single DNA molecules using rolling circle amplification. United States: N. p., 2013. Web.
Hutchison, III, Clyde A., & Smith, Hamilton O.. Amplification and cloning of single DNA molecules using rolling circle amplification. United States.
Hutchison, III, Clyde A., and Smith, Hamilton O.. Tue . "Amplification and cloning of single DNA molecules using rolling circle amplification". United States. https://www.osti.gov/servlets/purl/1531861.
@article{osti_1531861,
title = {Amplification and cloning of single DNA molecules using rolling circle amplification},
author = {Hutchison, III, Clyde A. and Smith, Hamilton O.},
abstractNote = {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.},
doi = {},
journal = {},
number = ,
volume = ,
place = {United States},
year = {2013},
month = {7}
}

Works referenced in this record:

TempliPhi, ⌽29 DNA Polymerase Based Rolling Circle Amplification of Templates for DNA Sequencing
journal, June 2002


Method for amplification of nucleic acids in solid media
patent, April 1997


Methods of amplifying and sequencing nucleic acids
patent-application, June 2005


Isothermal Strand-Displacement Amplification Applications for High-Throughput Genomics
journal, December 2002


Convergent DNA synthesis: a non-enzymatic dimerization approach to circular oligodeoxynucleotides
journal, January 1995


Rapid thermocycling for sample analysis
patent-application, October 2001


Rolling Circle DNA Synthesis:  Small Circular Oligonucleotides as Efficient Templates for DNA Polymerases
journal, January 1996


Cell-free cloning using φ29 DNA polymerase
journal, November 2005


A “Double Adaptor” Method for Improved Shotgun Library Construction
journal, April 1996


Rolling replication of short DNA circles.
journal, May 1995


Human parainfluenza virus-1 assay
patent, April 1998


Single-molecule PCR using water-in-oil emulsion
journal, April 2003


    Works referencing / citing this record:

    Individually synthesized primers to be used in whole genome amplification
    patent, March 2017