Premium
Characterization of acridone dyes for use in four‐decay detection in DNA sequencing
Author(s) -
Mihindukulasuriya Samila H.,
Morcone Tara K.,
McGown Linda B.
Publication year - 2003
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.200390017
Subject(s) - acridone , capillary electrophoresis , fluorescence , dna , chemistry , dna sequencing , primer (cosmetics) , photochemistry , chromatography , biochemistry , stereochemistry , physics , optics , organic chemistry
Four acridone dyes and dye‐labeled primers were characterized for use in four‐decay DNA sequencing. In the four‐decay scheme, fluorescence lifetime replaces spectral (“color”) selectivity for distinguishing between four base‐specific labels in a single‐lane capillary electrophoresis (CE) separation of the DNA fragments. Prior to the introduction of the acridone dyes, a major obstacle to four‐decay detection was the lack of four suitable dyes with resolvable lifetimes. The four acridone dyes, whether free in solution or tethered to DNA primer, exhibit significant differences among their lifetimes and are well‐suited to use together in four‐decay sequencing. The lifetimes of the four dye‐labeled DNA primers that were sequentially injected and detected on‐the‐fly in a 2% POP6 sequencing gel were 4, 6, 11 and 14 ns. A 405 nm violet laser diode provides optimal excitation of the four dyes.