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• W2028989854 endingPage "4351" @default.
• W2028989854 startingPage "4347" @default.
• W2028989854 abstract "Fluorescent dye-labeled DNA primers have been developed that exploit fluorescence energy transfer (ET) to optimize the absorption and emission properties of the label. These primers carry a fluorescein derivative at the 5' end as a common donor and other fluorescein and rhodamine derivatives attached to a modified thymidine residue within the primer sequence as acceptors. Adjustment of the donor-acceptor spacing through the placement of the modified thymidine in the primer sequence allowed generation of four primers, all having strong absorption at a common excitation wavelength (488 nm) and fluorescence emission maxima of 525, 555, 580, and 605 nm. The ET efficiency of these primers ranges from 65% to 97%, and they exhibit similar electrophoretic mobilities by gel electrophoresis. With argon-ion laser excitation, the fluorescence of the ET primers and of the DNA sequencing fragments generated with ET primers is 2- to 6-fold greater than that of the corresponding primers or fragments labeled with single dyes. The higher fluorescence intensity of the ET primers allows DNA sequencing with one-fourth of the DNA template typically required when using T7 DNA polymerase. With single-stranded M13mp18 DNA as the template, a typical sequencing reaction with ET primers on a commercial sequencer provided DNA sequences with 99.8% accuracy in the first 500 bases. ET primers should be generally useful in the development of other multiplex DNA sequencing and analysis methods." @default.
• W2028989854 created "2016-06-24" @default.
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• W2028989854 date "1995-05-09" @default.
• W2028989854 modified "2023-10-14" @default.
• W2028989854 title "Fluorescence energy transfer dye-labeled primers for DNA sequencing and analysis." @default.
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• W2028989854 doi "https://doi.org/10.1073/pnas.92.10.4347" @default.
• W2028989854 hasPubMedCentralId "https://www.ncbi.nlm.nih.gov/pmc/articles/41941" @default.
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