Fluorescence detection in automated DNA sequence analysis

Abstract

We have developed a method for the partial automation of DNA sequence analysis. Fluorescence detection of the DNA fragments is accomplished by means of a fluorophore covalently attached to the oligonucleotide primer used in enzymatic DNA sequence analysis. A different coloured fluorophore is used for each of the reactions specific for the bases A, C, G and T. The reaction mixtures are combined and co-electrophoresed down a single polyacrylamide gel tube, the separated fluorescent bands of DNA are detected near the bottom of the tube, and the sequence information is acquired directly by computer.

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Smith, L., Sanders, J., Kaiser, R. et al. Fluorescence detection in automated DNA sequence analysis. Nature 321, 674–679 (1986). https://doi.org/10.1038/321674a0

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