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

By F. Dean, John R. Nelson, T. Giesler, R. Lasken. Genome Research, 2001

1,081 citations recorded by the source corpus.

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.

Subfields: Cancer Research, Ecology, Molecular Biology

View Paper: https://www.semanticscholar.org/paper/029cdf4a42ffecc70b0f389538eb0a237d831cc0

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In Molecular Biology

1. Steven Lin
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Citations in the corpus

• Begomovirus disease complex: emerging threat to vegetable production systems of West and Central Africa (2015, methodology): “Recently, the most widely applied method for detecting and cloning begomoviral genomes is the non-sequencespecific approach that uses bacteriophage phi29 DNA polymerase [121,122], now referred to as RCA [123].”
• Real-time DNA sequencing from single polymerase molecules. (2010, background): “It is also very stable, maintaining constant enzymatic activities for up to several days (Dean et al., 2001; Nelson et al., 2002).”
• Real-Time DNA Sequencing from Single Polymerase Molecules (2009, background): “It is also very stable, maintaining constant enzymatic activities for up to several days (Dean et al., 2001; Nelson et al., 2002).”
• Somatic mutation in single human neurons tracks developmental and transcriptional history (2015, methodology): “Our data demonstrate that individual clones intermingle widely in the human cortex and that neurons from a given cortical region constitute Nuclei are isolated from a 0.5-cm 3 piece of frozen postmortem tissue by fluorescenceactivated nuclear sorting (FANS), and DNA is amplified by F29 polymerase–me”