Abstract
DNA consists of a continuous chain of deoxyribose units linked via phosphodiester bonds at their 5′ and 3′ positions and joined to one of the four nucleotide bases, Thymine (T), cytosine (C), the pyrimidines, guanine (G), or adenine (A), the purines, at their 1′ positions. It exists in organisms in the form of very long chains (5,000 base-pairs long, in the case of bacteriophage øX174, to 240,000,000 bp long, in the case of the largest human chromosome). Thus, in order to determine the sequence of a DNA molecule, the order of nucleotide bases along its sugar-phosphate backbone, it is first necessary to fragment the molecule into segments of a manageable size and to purify each fragment type. This is most readily accomplished by cloning into a plasmid or viral DNA vector. After amplification of the resulting hybrid DNA molecule, the cloned DNA segments (inserts) can be released for sequencing by cleavage with a restriction endonuclease.
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© 1984 Springer-Verlag Berlin Heidelberg
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Volckaert, G., Winter, G., Gaillard, C. (1984). DNA Sequencing. In: Pühler, A., Timmis, K.N. (eds) Advanced Molecular Genetics. Springer, Berlin, Heidelberg. https://doi.org/10.1007/978-3-642-69305-2_5
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DOI: https://doi.org/10.1007/978-3-642-69305-2_5
Publisher Name: Springer, Berlin, Heidelberg
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