What question did this study set out to answer?

To develop and characterize fluorescence energy transfer (ET) dye-labeled primers that optimize absorption and emission properties for sensitive DNA sequencing.

May 9, 1995Proceedings of the National Academy of SciencesOpen Access

Fluorescence energy transfer dye-labeled primers for DNA sequencing and analysis.

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Authors

JJJingfang JuStony Brook School CRC. RuanSoochow University CFCarl W. FullerEndocrine Research Solutions (United States)

Discussion

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Implication

Methodological study demonstrates enhanced fluorescence and 99.8% accuracy in DNA sequencing using energy-transfer primers, indicating potential to reduce template requirements in multiplex assays.

Key Points

To develop and characterize fluorescence energy transfer (ET) dye-labeled primers that optimize absorption and emission properties for sensitive DNA sequencing.
Synthesized four primers containing a 5'-fluorescein donor paired with distinct fluorescein or rhodamine acceptor dyes on modified thymidine residues.
Optimized donor-acceptor spacing for uniform excitation at 488 nm with distinct emission peaks at 525, 555, 580, and 605 nm.
Evaluated energy transfer efficiency, electrophoretic mobility, and sequencing accuracy using T7 DNA polymerase on single-stranded M13mp18 DNA templates.
ET primers exhibited energy transfer efficiencies between 65% and 97% and uniform electrophoretic mobilities.
Fluorescence intensity of ET primers and labeled sequencing fragments increased 2- to 6-fold compared to single-dye counterparts under argon-ion laser excitation.
Sequencing with ET primers reduced template requirements to one-fourth the standard amount and achieved 99.8% accuracy across the first 500 bases of M13mp18 DNA.

Cite This Study

Ju et al. (1995) studied this question.

synapsesocial.com/papers/6a0f440bb997c1a6759bc732 https://doi.org/10.1073/pnas.92.10.4347

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May 9, 1995Proceedings of the National Academy of SciencesOpen Access

Fluorescence energy transfer dye-labeled primers for DNA sequencing and analysis.

View Full Paper

Ask AI

Bookmark

Authors

JJJingfang JuStony Brook School CRC. RuanSoochow University CFCarl W. FullerEndocrine Research Solutions (United States)

Discussion

Member takes

Implication

Methodological study demonstrates enhanced fluorescence and 99.8% accuracy in DNA sequencing using energy-transfer primers, indicating potential to reduce template requirements in multiplex assays.

Key Points

To develop and characterize fluorescence energy transfer (ET) dye-labeled primers that optimize absorption and emission properties for sensitive DNA sequencing.
Synthesized four primers containing a 5'-fluorescein donor paired with distinct fluorescein or rhodamine acceptor dyes on modified thymidine residues.
Optimized donor-acceptor spacing for uniform excitation at 488 nm with distinct emission peaks at 525, 555, 580, and 605 nm.
Evaluated energy transfer efficiency, electrophoretic mobility, and sequencing accuracy using T7 DNA polymerase on single-stranded M13mp18 DNA templates.
ET primers exhibited energy transfer efficiencies between 65% and 97% and uniform electrophoretic mobilities.
Fluorescence intensity of ET primers and labeled sequencing fragments increased 2- to 6-fold compared to single-dye counterparts under argon-ion laser excitation.
Sequencing with ET primers reduced template requirements to one-fourth the standard amount and achieved 99.8% accuracy across the first 500 bases of M13mp18 DNA.

Cite This Study

Ju et al. (1995) studied this question.

synapsesocial.com/papers/6a0f440bb997c1a6759bc732 https://doi.org/10.1073/pnas.92.10.4347

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