Preparation and Direct Automated Cycle Sequencing of PCR Products

  • Susan E. Daniels
Part of the Methods in Molecular Biology™ book series (MIMB, volume 65)


The polymerase chain reaction (PCR) is well known for being a rapid and versatile method for the amplification of defined target DNA sequences. This technique can be applied to a variety of research areas, such as the identification and typing of single nucleotide substitutions of DNA sequence polymorphisms, and genetic mapping (1, 2, 3, 4).


Polymerase Chain Reaction Product Cycle Sequencing Microfuge Tube Single Nucleotide Substitution Chain Terminator 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.


  1. 1.
    Orita, M., Suzuki, Y., Sekeiya, T., and Hayashi, K. (1989) Rapid and sensitive detection of point mutations and DNA polymorphisms using the polymerase chain reaction. Genomics 5, 874–879.PubMedCrossRefGoogle Scholar
  2. 2.
    Kwok, P. Y., Carlson, C., Yager, T. D., Ankener, W., and Nickerson, D. A. (1994) Comparative analysis of human DNA variations by fluorescence based sequencing of PCR products. Genomics 23, 138–144.PubMedCrossRefGoogle Scholar
  3. 3.
    Martin-Gallardo, A., McCombie, W. R., Gocayne, J. D., Fitzgerald, M. G., Wallace, S., Lee, B. M. B., Lamerdin, J., Trapp, S., Kelly, J. M., Liu, L. I., Dubrick, M., Johnston-Dow, L. A., Kerlavage, A. R., de Jong, P., Carrano, A., Fields, C., and Venter, J. C. (1992) Automated DNA sequencing and analysts of 106 kilobases from human chromosome 19q 13.3. Nature Genetics 1, 34–39.PubMedCrossRefGoogle Scholar
  4. 4.
    NIH/CEPH Collaborative Mapping Group (1992) A comprehensive genetic linkage map of the human genome. Science 258, 67–86.CrossRefGoogle Scholar
  5. 5.
    Mullis, K. B. and Faloona, F. A. (1987) Specific synthesis of DNA in vitro via a polymerase catalysed chain reaction. Methods Enzymol. 155, 335–350.PubMedCrossRefGoogle Scholar
  6. 6.
    Sanger, F., Nicklen, S., and Coulson, A. R. (1977) DNA sequencing with chain terminating inhibitors. Proc. Natl. Acad. Sci. USA 74, 5463–5467.PubMedCrossRefGoogle Scholar
  7. 7.
    Maxam, A. M. and Gilbert, W. (1977) A new method for sequencing DNA. Proc. Natl. Acad. Sci. USA 14, 560–564.CrossRefGoogle Scholar
  8. 8.
    Carothers, A. M., Urlaub, G., Mucha, J., Grunberger, D., and Chasin, L. A. (1989) Point mutation analysis in a mammalian gene: rapid preparation of total RNA, PCR amplification of cDNA, and Tag sequencing by a novel method. Biotechniques 7, 494–499.PubMedGoogle Scholar
  9. 9.
    McBride, L. J., Koepf, S. M., Gibbs, R. A., Nyugen, P., Salser, W., Mayrand, P. E., Hunkapiller, M. W., and Kronick, M. N. (1989) Automated DNA sequencing methods involving polymerase chain reaction. Clin. Chem. 35, 2196–2201.PubMedGoogle Scholar
  10. 10.
    Tracy, T. E. and Mulcahy, L. S. (1991) A simple method for direct automated sequencing of PCR fragments. Biotechniques 11(1), 68–75.PubMedGoogle Scholar
  11. 11.
    Rosenthal, A. and Charnock Jones, D. S. (1992) New protocols for DNA sequencing with dye terminators. DNA Sequence 3, 61–64.PubMedGoogle Scholar
  12. 12.
    Kelley, J. M. (1994) Automated dye terminator DNA sequencing, in Automated DNA Sequencing and Analysis (Adams, M. A., Fields, C., and Venter, J. C., eds.), Academic, London, pp. 175–181.Google Scholar
  13. 13.
    Hawkins, T. L. and Sulston, J. E. (1991) The resolution of compressions in automated fluorescent sequencing. Nucleic Acids Res. 19(10), 2784.PubMedCrossRefGoogle Scholar

Copyright information

© Humana Press Inc., Totowa, NJ 1996

Authors and Affiliations

  • Susan E. Daniels
    • 1
  1. 1.Wellcome Trust Centre for Human GeneticsOxfordUK

Personalised recommendations