PMID: 8604350Mar 15, 1996Paper

Cassette labeling for facile construction of energy transfer fluorescent primers

Nucleic Acids Research
J JuR A Mathies

Abstract

DNA primer sets, labeled with two fluorescent dyes to exploit fluorescence energy transfer (ET), can be efficiently excited with a single laser line and emit strong fluorescence at distinctive wavelengths. Such ET primers are superior to single fluorophore-labeled primers for DNA sequencing and other multiple color-based analyses [J. Ju, C. Ruan, C. W. Fuller, A. N. Glazer and R. A. Mathies (1995) Proc. Natl. Acad. Sci. USA 92, 4347-4351]. We describe here a novel method of constructing fluorescent primers using a universal ET cassette that can be incorporated by conventional synthesis at the 5'-end of an oligonucleotide primer of any sequence. In this cassette, the donor and acceptor fluorophores are separated by a polymer spacer (S6) formed by six 1',2'-dideoxyribose phosphate monomers (S). The donor is attached to the 5' side of the ribose spacer and the acceptor to a modified thymidine attached to the 3' end of the ribose spacer in the ET cassette. The resulting primers, labeled with 6-carboxy-fluorescein as the donor and other fluorescein and rhodamine dyes as acceptors, display well-separated acceptor emission spectra with 2-12-fold enhanced fluorescence intensity relative to that of the corresponding single dye-labeled p...Continue Reading

References

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Citations

Apr 15, 2005·Mutation Research·John R EdwardsJingyue Ju
Feb 13, 2003·Journal of Neuroscience Methods·Dmitri ProudnikovMary Jeanne Kreek
Feb 1, 1997·Current Opinion in Biotechnology·A N Glazer, R A Mathies
Mar 21, 1998·Current Opinion in Biotechnology·S Beck, P Sterk
Mar 20, 2012·Chemical Reviews·Yin Nah Teo, Eric T Kool
Jul 15, 1997·Nucleic Acids Research·L G LeeJ M Cassel
Feb 12, 1998·Nucleic Acids Research·B B RosenblumS M Chen
Nov 3, 2001·Nucleic Acids Research·J R EdwardsJ Ju
Oct 6, 1998·Annual Review of Biochemistry·S Verma, F Eckstein
May 18, 2001·Analytical Biochemistry·L BertiR A Mathies
Jan 8, 2003·Proceedings of the National Academy of Sciences of the United States of America·Zengmin LiJingyue Ju
Apr 15, 2005·Proceedings of the National Academy of Sciences of the United States of America·Hameer RuparelJingyue Ju
Nov 27, 2004·Electrophoresis·Cheuk-Wai KanAnnelise E Barron
Sep 21, 2001·Nucleosides, Nucleotides & Nucleic Acids·T S RaoS Kumar
Nov 25, 2003·Nature Reviews. Genetics·Sobin KimJingyue Ju
Apr 23, 2008·Current Protocols in Nucleic Acid Chemistry·N T Thuong, U Asseline
Nov 2, 2007·Chemical Reviews·Karel Kleparník, Petr Bocek

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