DNA Sequencing by the Chemical Method
- Eran Pichersky
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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. 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).
- Maxam, A. M. and Gilbert, W. (1980) Sequencing end-labeled DNA with base-specific chemical cleavages. Methods Enzymol. 65, 499–560. CrossRef
- Sanger, F., Nicklen, S., and Coulson, A. R. (1977) DNA sequencing with chain-terminating inhibitors. Proc. Natl. Acad. Sci USA 74, 5463–5467. CrossRef
- Maniatis, T., Fritsch, E. F., and Sambrook, J. (1982) Molecular Cloning, A Laboratory Manual. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY.
- DNA Sequencing by the Chemical Method
- Book Title
- PCR Sequencing Protocols
- pp 133-136
- Print ISBN
- Online ISBN
- Series Title
- Methods in Molecular Biology™
- Series Volume
- Series ISSN
- Springer New York
- Copyright Holder
- Humana Press
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