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      Rapid amplification of plasmid and phage DNA using Phi 29 DNA polymerase and multiply-primed rolling circle amplification.

      1 , , ,
      Genome research
      Cold Spring Harbor Laboratory

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          Abstract

          We describe a simple method of using rolling circle amplification to amplify vector DNA such as M13 or plasmid DNA from single colonies or plaques. Using random primers and phi29 DNA polymerase, circular DNA templates can be amplified 10,000-fold in a few hours. This procedure removes the need for lengthy growth periods and traditional DNA isolation methods. Reaction products can be used directly for DNA sequencing after phosphatase treatment to inactivate unincorporated nucleotides. Amplified products can also be used for in vitro cloning, library construction, and other molecular biology applications.

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          Author and article information

          Journal
          Genome Res
          Genome research
          Cold Spring Harbor Laboratory
          1088-9051
          1088-9051
          Jun 2001
          : 11
          : 6
          Affiliations
          [1 ] Molecular Staging, Inc., New Haven, Connecticut 06511, USA.
          Article
          10.1101/gr.180501
          311129
          11381035
          02862fbe-a178-4b93-ac28-1ca74c7299ba
          History

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