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Related Experiment Videos

New energy transfer dyes for DNA sequencing

L G Lee1, S L Spurgeon, C R Heiner

  • 1PE Applied Biosystems, 850 Lincoln Centre Drive, Foster City, CA 94404, USA. leelg@perkin-elmer.com

Nucleic Acids Research
|July 15, 1997
PubMed
Summary

New energy transfer dyes enhance automated DNA sequencing. These novel dyes offer improved brightness and reduced noise, leading to a 4- to 5-fold increase in signal-to-noise ratio for more accurate sequencing results.

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Area of Science:

  • Biochemistry
  • Molecular Biology
  • Analytical Chemistry

Background:

  • Automated DNA sequencing relies on fluorescent dyes for fragment detection.
  • Existing dye systems can suffer from spectral overlap and noise, limiting accuracy.
  • Energy transfer dyes offer potential for improved signal and reduced noise.

Purpose of the Study:

  • To synthesize and evaluate novel energy transfer dyes for automated DNA sequencing.
  • To improve signal-to-noise ratio and reduce multicomponent noise in DNA sequencing.
  • To demonstrate the utility of these dyes in sequencing large DNA fragments.

Main Methods:

  • Synthesis of four energy transfer dyes using fluorescein donors and novel dichlororhodamine acceptors.
  • Utilized a rigid amino acid linker (4-aminomethylbenzoic acid) for dye separation.

Related Experiment Videos

  • Tested dye performance in an automated sequencing instrument with a dual-line argon ion laser.

    • Main Results:

    • The new energy transfer dyes exhibited 2-2.5 times greater brightness compared to standard dye-primers.
    • A 2-fold reduction in multicomponent noise was observed.
    • Overall signal-to-noise ratio improved by 4- to 5-fold, demonstrated by sequencing an 80 kb BAC DNA insert.
    • Dichlororhodamine components showed a 20-25% reduction in extinction coefficients and quantum yields.

    Conclusions:

    • The developed energy transfer dyes significantly enhance automated DNA sequencing performance.
    • The dyes provide a substantial improvement in signal-to-noise ratio and reduction in noise.
    • These novel dyes are effective for sequencing large DNA fragments, offering a valuable tool for molecular biology research.