Abstract
Fluorescent cycle sequencing of PCR products is a multistage process and several methodologies are available to perform each stage. This chapter will describe the more commonly utilised dye-terminator cycle sequencing approach using BigDye® terminator chemistry (Applied Biosystems) ready for analysis on a 3730 DNA genetic analyzer. Even though DNA sequencing is one of the most common and robust techniques performed in molecular laboratories it may not always produce desirable results. The causes of the most common problems will also be discussed in this chapter.
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References
Sanger, F., Nicklen, S., and Coulson, A. R. (1977) DNA sequencing with chain terminating inhibitors. Proc. Natl Acad. Sci. USA 74, 5463–5467.
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© 2011 Humana Press
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Wallis, Y., Morrell, N. (2011). Automated DNA Sequencing. In: Theophilus, B., Rapley, R. (eds) PCR Mutation Detection Protocols. Methods in Molecular Biology, vol 688. Humana Press. https://doi.org/10.1007/978-1-60761-947-5_12
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DOI: https://doi.org/10.1007/978-1-60761-947-5_12
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Publisher Name: Humana Press
Print ISBN: 978-1-60761-946-8
Online ISBN: 978-1-60761-947-5
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