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Amplification with Arbitrary Primers

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Part of the book series: Methods in Molecular Biology™ ((MIMB,volume 54))

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Abstract

Several methods have been published that rely on the use of short oligonucleotide primers with arbitrary sequences to amplify discrete DNA fragments by the polymerase chain reaction (PCR) (1,2). Typically, a single arbitrary primer is used in each reaction and amplification is achieved when the same sequence is present in inverted orientation at two sites separated by less than a few kilobases. These methods have several advantages:

  1. 1.

    They can be used to produce quickly large numbers of discrete DNA fragments without prior sequence information;

  2. 2.

    Owing to the random nature of the process, the amplified fragments are likely to be evenly distributed across a genomic region;

  3. 3.

    Because they do not rely on the presence of species-specific repetitive sequences (e.g., Alu repeats [3]), they can be used to analyze the genome of any species.

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References

  1. Williams, J. G. K., Kubelik, A. R., Livak, K. J., Rafalski, J. A., and Tingey, S. V. (1990) DNA polymorphisms amplified by arbitrary primers are useful genetic markers. Nucleic Acids Res. 18, 6531–6535.

    Article  PubMed  CAS  Google Scholar 

  2. Welsh, J. and McClelland, M. (1990) Fingerprinting genomes using PCR with arbitrary primers. Nucleic Acids Res. 18, 7213–7218.

    Article  PubMed  CAS  Google Scholar 

  3. Nelson, D. L., Ledbetter, S. A., Corbo, L., Victoria, M. F., Ramirez-Solis, R., Webster, T., et al. (1989) Alu polymerase chain reaction: a method for rapid isolation of human-specific sequences from complex DNA sources. Proc. Natl. Acad. Sci. USA 86, 6686–6690.

    Article  PubMed  CAS  Google Scholar 

  4. Di Rienzo, A., Peterson, A., Das, S., and Freimer, N. B. (1993) Genome mapping by arbitrary amplification of yeast artificial chromosomes. Mammalian Genome 4, 359–363.

    Article  PubMed  Google Scholar 

  5. Burmeister, M., diSibio, G., Cox, D. R., and Myers, R. M. (1991) Identification of polymorphisms by genomic denaturing gradient gel electrophoresis: application to the proximal region of human chromosome 21. Nucleic Acids Res. 19, 1475–1481.

    Article  PubMed  CAS  Google Scholar 

  6. Olson, M., Hood, L., Cantor, C., and Botstein, D. (1989) A common language for physical mapping of the human genome. Science 245, 1434–1435.

    Article  PubMed  CAS  Google Scholar 

  7. Feinberg, A. P. and Vogelstein, B. (1984) Addendum: a technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal. Biochem. 137, 266,267

    Article  PubMed  CAS  Google Scholar 

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© 1996 Humana Press Inc.

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Di Rienzo, A., Peterson, A.C., Freimer, N.B. (1996). Amplification with Arbitrary Primers. In: Markie, D. (eds) YAC Protocols. Methods in Molecular Biology™, vol 54. Humana Press. https://doi.org/10.1385/0-89603-313-9:123

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  • DOI: https://doi.org/10.1385/0-89603-313-9:123

  • Publisher Name: Humana Press

  • Print ISBN: 978-0-89603-313-9

  • Online ISBN: 978-1-59259-541-9

  • eBook Packages: Springer Protocols

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