Abstract
The dideoxy chain-termination method of DNA sequence analysis involves the synthesis of a DNA strand from a single-stranded template (1). The enzymatic synthesis is initiated at the site where an oligonucleotide primer anneals to the template. The most consistently satisfactory sequencing results are probably obtained by using bacteriophage M13 (2,3) or phagemid (4) single-stranded DNA as the template. However, plasmid DNA that has been denatured can also serve as satisfactory template DNA. It is particularly important when preparing plasmid DNA for sequencing to give the utmost care and attention to the DNA isolation and purification techniques. Most problems that occur with plasmid sequencing are related to poor quality template.
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© 1993 Humana Press Inc. Totowa, New Jersey
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Griffin, H.G., Griffin, A.M. (1993). Plasmid Sequencing. In: Griffin, H.G., Griffin, A.M. (eds) DNA Sequencing Protocols. Methods in Molecular Biology™, vol 23. Humana Press. https://doi.org/10.1385/0-89603-248-5:131
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DOI: https://doi.org/10.1385/0-89603-248-5:131
Publisher Name: Humana Press
Print ISBN: 978-0-89603-248-4
Online ISBN: 978-1-59259-510-5
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