DNA Sequencing

  • Hugh G. Griffin
  • Annette M. Griffin
Part of the Methods in Molecular Biology™ book series (MIMB, volume 23)

Abstract

Methods to determine the sequence of DNA were developed in the late 1970s (1,2) and have revolutionized the science of molecular genetics. The DNA sequences of many different genes from diverse sources have been determined, and the information is stored in international databanks such as EMBL, GenBank, and DDBJ. Many scientists now accept that sequence analysis will provide an increasingly useful approach to the characterization of biological systems. Projects are already underway to map and sequence the entire genome of organisms such as Escherichia coli, Saccharomyces cerevisiae, Caenorhabditis elegans, and Homo sapiens. In the recent past, large-scale sequencing projects such as these were often dismissed as prohibitively expensive and of little short-term benefit, while DNA sequencing itself was seen as a repetitive and unintellectual pursuit. However, this view is now changing and most scientists recognize the importance of DNA sequence data and perceive DNA sequencing as a valuable and often indispensable aspect of their work. Recent technological advances, especially in the area of automated sequencing, have removed much of the drudgery that used to be associated with the technique, and modern innovative computer software has greatly simplified the analysis and manipulation of sequence data. Large-scale sequencing projects, such as the Human Genome Project, produce the DNA sequences of many unknown genes. Such data provide an impetus for molecular biologists to apply the techniques of reverse genetics to produce probes and antibodies that can be used to identify the gene product, its cellular location, and its time of appearance in the developing cell (3). A function can be assigned by mutant analysis or by comparison of the deduced amino acid sequence with proteins of known function. Therefore, DNA sequencing can act as a catalyst to stimulate future research into many diverse areas of science.

Keywords

Hydroxyl Electrophoresis Polyacrylamide Saccharomyces dNTP 

References

  1. 1.
    Sanger, F., Nicklen, S., and Coulson, A. R. (1977) DNA sequencing wrth chaintermmator inhibitors. Proc. Natl. Acad. Sci USA 74, 5463–5467.PubMedCrossRefGoogle Scholar
  2. 2.
    Maxam, A. M and Gilbert, W (1977) A new method for sequencing DNA Proc. Natl. Acad. Scl USA 74, 560–564.CrossRefGoogle Scholar
  3. 3.
    Barrell, B. (1991) DNA sequencmg: present limitations and prospects for the future. FASEB J 5, 40–45PubMedGoogle Scholar
  4. 4.
    Sambrook, J., Fritsch, E F., and Maniatrs, T. (1989) Molecular Cloning. A Laboratory Manual 2d ed, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY.Google Scholar
  5. 5.
    Maxam, A. M. and Gilbert, W. (1980) Sequencing end-labeled DNA with basespecific chemical cleavages. Meth. Enzym. 65, 499–560.PubMedCrossRefGoogle Scholar
  6. 6.
    Ambrose, B J. B. and Pless, R. C. (1987) DNA sequencing. Chemical methods. Meth. Enzymol. 152, 522–538.PubMedCrossRefGoogle Scholar
  7. 7.
    Volckaert, G. (1987) A systematic approach to chemical DNA sequencing by subcloning in pGV451 and derrved vectors. Meth Enzym. 155, 231–250PubMedCrossRefGoogle Scholar
  8. 8.
    Eckert, R. L. (1987) New vectors for rapid sequencing of DNA fragments by chemical degradation. Gene 51, 247–254.PubMedCrossRefGoogle Scholar
  9. 9.
    Dolz, R. (1993) Fragment assembly programs, in DNA sequencmg: Computer Analysis of Sequence Data, (Griffin, A. M. and Grrffm, H. G., eds.), Humana Press, Totowa, NJ. (Ch. 2).Google Scholar
  10. 10.
    Staden, R. (1992) Managing sequencing projects, in DNA sequencing. Computer Analysis of Sequence Data, (Griffin, A. M. and Griffin, H. G., eds.), Humana Press, Totowa, NJ. (Ch 17).Google Scholar
  11. 11.
    Hunkapiller, T, Karser, R J., Kopp, B F, and Hood, L. (1991) Large-scale and automated DNA sequence determination. Science 254, 59–67PubMedCrossRefGoogle Scholar

Copyright information

© Humana Press Inc. Totowa, New Jersey 1993

Authors and Affiliations

  • Hugh G. Griffin
    • 1
  • Annette M. Griffin
    • 1
  1. 1.Institute of Food ResearchNorwichEngland

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