Abstract
The chemical method of sequencing DNA (1) has some advantages and some disadvantages compared with the enzymatic method (2). The major disadvantage is that it takes more time to produce the same amount of sequence. This is so for two main reasons. First, the DNA has to be end-labeled and then reisolated prior to the actual chemical sequencing reactions, a process that usually requires an additional day (see Chapter 31). Also, because more DNA is used in the reaction and because the lower specific activity of the sequenced DNA requires the use of an intensifying screen in the autoradiography, bands are not as sharp as in the enzymatic method and therefore it is difficult to obtain reliable sequence past about nucleotide 250 (unless very long gels are run).
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References
Maxam, A. M and Gilbert, W. (1980) Sequencing end-labeled DNA with basespecific chemical cleavages Meth Enzymol. 65, 499–560
Sanger, F., Nicklen, S., and Coulson, A R. (1977) DNA sequencing with chamterminating inhibitors Proc Natl. Acad. Sci USA 74, 5463–5467.
Maniatis, T., Fritsch, E. F., and Sambrook, J (1982) Molecular Cloning, A Laboratory Manual. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY
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© 1993 Humana Press Inc. Totowa, New Jersey
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Pichersky, E. (1993). DNA Sequencing the Chemical Method. In: Griffin, H.G., Griffin, A.M. (eds) DNA Sequencing Protocols. Methods in Molecular Biology™, vol 23. Humana Press. https://doi.org/10.1385/0-89603-248-5:255
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DOI: https://doi.org/10.1385/0-89603-248-5:255
Publisher Name: Humana Press
Print ISBN: 978-0-89603-248-4
Online ISBN: 978-1-59259-510-5
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