Fluorescence detection in automated DNA sequence analysis

  title={Fluorescence detection in automated DNA sequence analysis},
  author={Lloyd M. Smith and Jane Z. Sanders and Robert J. Kaiser and Peter Hughes and Chris Dodd and Charles R. Connell and Cheryl R. Heiner and Stephen B. H. Kent and Leroy E. Hood},
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… 

Rapid DNA sequencing based upon single molecule detection.

Colorimetric-detected DNA sequencing.

  • S. Beck
  • Biology, Chemistry
    Analytical biochemistry
  • 1987

A system for rapid DNA sequencing with fluorescent chain-terminating dideoxynucleotides.

A DNA sequencing system based on the use of a novel set of four chain-terminating dideoxynucleotides, each carrying a different chemically tuned succinylfluorescein dye distinguished by its fluorescent emission, and the sequence analysis of M13 phage DNA made with this system is described.


DNA sequence analysis is one of the most important techniques of modern molecular biology. It permits the rapid and complete determination of the precise chemical structure of the genetic material.

A ligase-mediated gene detection technique.

An assay for the presence of given DNA sequences has been developed, based on the ability of two oligonucleotides to anneal immediately adjacent to each other on a complementary target DNA molecule, which permits the rapid and standardized identification of single-copy gene sequences in genomic DNA.

High-speed DNA sequencing: an approach based upon fluorescence detection of single molecules.

A laser based technique for the rapid sequencing of large fragments of DNA based upon the detection of single, fluorescently tagged nucleotides cleaved from a single DNA fragment which is orders of magnitude faster than existing methods.



DNA sequencing with chain-terminating inhibitors.

A new method for determining nucleotide sequences in DNA is described, which makes use of the 2',3'-dideoxy and arabinon nucleoside analogues of the normal deoxynucleoside triphosphates, which act as specific chain-terminating inhibitors of DNA polymerase.

DNA sequence analysis by primed synthesis.

  • A. Smith
  • Biology, Chemistry
    Methods in enzymology
  • 1980

The synthesis of oligonucleotides containing an aliphatic amino group at the 5' terminus: synthesis of fluorescent DNA primers for use in DNA sequence analysis.

A rapid and versatile method has been developed for the synthesis of oligonucleotides which contain an aliphatic amino group at their 5' terminus. This amino group reacts specifically with a variety

Non-radioactive hybridization probes prepared by the chemical labelling of DNA and RNA with a novel reagent, photobiotin.

The sensitivity of detection of target RNA in dot-blots and Northern blots was equivalent to that obtained with 32p-labelled DNA probes, and photobiotin was also used for the labelling of proteins with biotin.

Solid-phase methods for sequencing of nucleic acids I. Simultaneous sequencing of different oligodeoxyribonucleotides using a new, mechanically stable anion-exchange paper.

A solid-phase method for simultaneous sequencing of large numbers of oligodeoxyribonucleotides has been developed using a new, mechanically stable anion-exchange paper. The excellent mechanical

Oligonucleotide synthesis : a practical approach

An introduction to modern methods of DNA synthesis and chromatographic purification of synthetic oligonucleotides is presented.