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Rapid Amplification of Plasmid and Phage DNA Using Phi29 DNA Polymerase and Multiply-Primed Rolling Circle Amplification
Open accessAbstract
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 φ29 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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Citations
Cited by
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Milani, C., Duranti, S., Bottacini, F., Casey, E., Turroni, F., Mahony, J., Belzer, C., Delgado Palacio, S., Arboleya Montes, S., Mancabelli, L., et al. (2017) · Microbiology and Molecular Biology Reviews · doi:10.1128/mmbr.00036-17 · Cited by 1,950
Comprehensive human genome amplification using multiple displacement amplification
Dean, F. B., Hosono, S., Fang, L., Wu, X., Faruqi, A. F., Bray-Ward, P., Sun, Z., Zong, Q., Du, Y., Du, J., et al. (2002) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.082089499 · Cited by 1,467
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Metadata
- Authors
- Dean, F. B., Nelson, J. R., Giesler, T. L., & Lasken, R. S.
- Venue
- Genome Research
- Publisher
- Cold Spring Harbor Laboratory
- Published
- 2001-06-01
- Volume
- 11
- Issue
- 6
- Pages
- 1095-1099
- ISSN
- 1088-9051 (ISSN-L)
- 1549-5469
- OpenAlex
- W1968762427
- Cited by
- 1,067
- References
- 0
- Type
- posted content
- Language
- en
- Open access
- bronze
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- Dean, F. B., Nelson, J. R., Giesler, T. L., & Lasken, R. S. (2001). Rapid Amplification of Plasmid and Phage DNA Using Phi29 DNA Polymerase and Multiply-Primed Rolling Circle Amplification. Genome Research. https://doi.org/10.1101/gr.180501
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