Abstract
Knowledge of the DNA primary sequence leads to a closer understanding of the structural organization of DNA, the deduction of an amino acid sequence and allows for the analysis of (clinically relevant) mutations. The first sequencing methods were quite labor-intensive and time-consuming (Wu and Taylor 1971; Robertson et al. 1973; Sanger et al. 1973; Ziff et al. 1973). However, two rapid and universally applicable methods have emerged: Maxam and Gilbert used chemical agents producing defined fragments of DNA (Maxam and Gilbert 1977). These fragments are made visible by Polyacrylamide gel electrophoresis using radioactive markers. The original DNA sequence can be deduced from the length of fragments formed. The second technique has been developed by Sanger in 1977 (Sanger et al. 1977). This method, the chain termination method, is most widely used nowadays, and is based on enzymatic steps (Davis et al. 1986; Sambrook et al. 1989; Ausubel et al. 1990).
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© 1999 Springer-Verlag Berlin Heidelberg
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Deichmann, K. (1999). Sequencing. In: Hildebrandt, F., Igarashi, P. (eds) Techniques in Molecular Medicine. Springer Lab Manual. Springer, Berlin, Heidelberg. https://doi.org/10.1007/978-3-642-59811-1_13
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DOI: https://doi.org/10.1007/978-3-642-59811-1_13
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