Time-resolved laser-induced fluorescence study on dyes used in DNA sequencing

Journal Article · Fri Jan 01 00:00:00 EST 1993 · Applied Spectroscopy; (United States)

DOI:https://doi.org/10.1366/0003702934048442· OSTI ID:6951506

Chang, Kaisyang; Force, R K ^[1]

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1. Univ. of Rhode Island, Kingston (United States)

Research on the time-resolved fluorescence of fluorescein isothiocyanate, NBD, tetramethylrhodamine isothiocyanate, and Texas Red - the dyes used for fluorescence-based DNA sequencing - is described. Mean fluorescence lifetiems in both aqueous buffer solution and 5.3%T, 4.8%C polyacrylamide gel were determined as a function of excitation wave-lengths at 337, 470, and 550 nm and were found to be 3.5, 1.1, 2.5, and 4.3 ns; the detection limits are 10, 200, 200 and 200 amol for FITC, NBD, TEMR, and T. Red, respectively. Comparisons of fluorescence parameters between the conjugated dyes and the free dyes are also reported. Results on the optimization of the excitation source wavelengths to improve sensitivity and reduce background scattering in polyacrylamide gel are also reported. Time-resolved fluorescence was successfully applied to resolve spectral overlapping of emissions in both solution and in polyacrylamide gel. 12 refs., 6 figs., 1 tab.

• OSTI ID: 6951506
• Journal Information: Applied Spectroscopy; (United States), Vol. 47:1; ISSN 0003-7028
• Country of Publication: United States
• Language: English

Related Subjects

37 INORGANIC
ORGANIC
PHYSICAL AND ANALYTICAL CHEMISTRY
59 BASIC BIOLOGICAL SCIENCES
DYES
FLUORESCENCE SPECTROSCOPY
AQUEOUS SOLUTIONS
DNA SEQUENCING
ELECTROPHORESIS
FLUORESCEIN
GELS
LASER SPECTROSCOPY
TIME RESOLUTION
AROMATICS
CARBOXYLIC ACIDS
COLLOIDS
DISPERSIONS
EMISSION SPECTROSCOPY
HYDROXY ACIDS
HYDROXY COMPOUNDS
MIXTURES
ORGANIC ACIDS
ORGANIC COMPOUNDS
PHENOLS
POLYPHENOLS
RESOLUTION
SOLUTIONS
SPECTROSCOPY
STRUCTURAL CHEMICAL ANALYSIS
TIMING PROPERTIES
400102* - Chemical & Spectral Procedures
550400 - Genetics