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A three-wavelength labeling approach for DNA sequencing using energy transfer primers and capillary electrophoresis



A three-wavelength labeling approach for DNA sequencing using energy transfer primers and capillary electrophoresis



Electrophoresis 19(8-9): 1403-1414



Capillary electrophoresis DNA sequencing has been accomplished by using four different energy transfer primers and three fluorescence detection channels. Methods have also been developed to deconvolve the three-color data into the four base concentrations. The nonnegative least squares and model selection method resulted in the best accuracy. The three-color data were compared to sequencing data obtained using four detection channels and four energy transfer primers. The average accuracy rates obtained over three 500 base M13mp18 runs using three-color coding were 96% including 18 uncallable compressions and 99.6% if these compressions are excluded. The average accuracy rate obtained using four-color coding was 96.3% including 18 uncallable compressions and 99.9% if these compressions are excluded. Although it is unlikely that three-color schemes will replace four-color sequencing, these methods have exposed basic concepts that will be useful in the development of higher-order multiplex coding methods for DNA analysis.

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Accession: 008084245

Download citation: RISBibTeXText

PMID: 9694290

DOI: 10.1002/elps.1150190835



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