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In the used strategy for automated fluorescent sequencing, fluorescent dye labels are incorporated into DNA extension products using 5’-dye labelled primers (dye primers) or 3’-dye labelled dideoxynucleotide triphosphates (dye terminators). Both methods were used in this thesis.
The used PE Applied Biosystems DNA sequencer (ABI 377) detects fluorescence from four different dyes that are used to identify the A, C, G, and T extension reactions. Each dye emits light at a different wavelength when excited by an argon ion laser. All four colours and therefore all four bases can be detected and distinguished in a single lane (Figure 2.2).
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© 2001 Alexander Binder
