Biopolym. Cell. 1995; 11(3-4):35-41.
Preparation of the fluorescein reagent for solid-phase oligonucleotide 5'-labelling and its use for the synthesis of fluorescently labelled PCR primers for HIV-1 detection
1Dubey I. Ya., 1Fedoryak O. D., 1Tkachak O. M., 1Fedoryak D. M.
  1. Institute of Bioorganic Chemistry and Petrochemistry, NAS of Ukraine
    1, Murmans'ka Str., Kyiv, Ukraine, 02094


The preparative synthesis of fluorescein H-phosphonate reagent for solid phase 'labelling of oligonucteotides is described. Fluorescein-labelted thymidine and two PCR primers for HIV-1 detection were synthesized using this reagent.


[1] Mundy CR, Cunningham MW, Read CA. Nucleic acids labelling and detection. Essential mol. biol. Ed. TA. Brown. Oxford : IRL, 1991. Vol. 2: 57-109.
[2] Kessler C. General aspects of nonradioactive labelling and detection. Nonradio­ active labelling and detection of biomol. Penzberg, 1992: 1-24.
[3] Koller E. Fluorescent labels for use in biology and biomedicine. Appl Fluoresc Technol. 1989; 1(2):1-8.
[4] Narayanan S. Overview of principles and current uses of DNA probes in clinical and laboratory medicine. Ann Clin Lab Sci. 1992;22(6):353-76.
[5] DNA probes. Eds G. H. Keller, M. M. Manak. MacMillan press, 1993. 656 p.
[6] Ansorge W, Sproat B, Stegemann J, Schwager C, Zenke M. Automated DNA sequencing: ultrasensitive detection of fluorescent bands during electrophoresis. Nucleic Acids Res. 1987;15(11):4593-602.
[7] Loke SL, Stein CA, Zhang XH, Mori K, Nakanishi M, Subasinghe C, Cohen JS, Neckers LM. Characterization of oligonucleotide transport into living cells. Proc Natl Acad Sci U S A. 1989;86(10):3474-8.
[8] Oligonucleotide synthesis a practical approach. Ed. M. J. Gait. Oxford : ORL press, 1984. 218 p.
[9] Kalashnikov VV, Samukov VV, Shubina TN, Yamshchikov VF. Application of the reverse-phase chromatography in oligonucleotide synthesis. Russian Journal of Bioorganic Chemistry 1983; 9(5):666-72
[10] Froehler BC, Ng PG, Matteucci MD. Synthesis of DNA via deoxynucleoside H-phosphonate intermediates. Nucleic Acids Res. 1986;14(13):5399-407.
[11] Dubey IYa, Ljapina TV, Galkin AP, Fedorjak DM. Obtaining of the highly efficient polymer support for the synthesis of the DNA fragments on the basis on the microspheric silica «Silochrom-2». Biopolym Cell. 1993; 9(4):26-31.
[12] Goodchild J. Conjugates of oligonucleotides and modified oligonucleotides: a review of their synthesis and properties. Bioconjug Chem. 1990;1(3):165-87.
[13] Korshun VA, Berlin YuA. Introduction of nonradioactive reporter groups into synthetic oligonucleotides and their detection. Russian Journal of Bioorganic Chemistry 1994; 20(6):565-616
[14] Beaucage SL, Iyer RP. The Functionalization of Oligonucleotides Via Phosphoramidite Derivatives. Tetrahedron. 1993;49(10):1925–63.
[15] Giusti WG, Adriano T. Synthesis and characterization of 5'-fluorescent-dye-labeled oligonucleotides. PCR Methods Appl. 1993;2(3):223-7.
[16] Haughland RP. Handbook of fluorescent probes and research chemicals. Junction City: Mol. Probes Inc., 1985. 124 p.
[17] Agrawal S, Christodoulou C, Gait MJ. Efficient methods for attaching non-radioactive labels to the 5' ends of synthetic oligodeoxyribonucleotides. Nucleic Acids Res. 1986;14(15):6227-45.
[18] Schubert F, Ahlert K, Cech D, Rosenthal A. One-step labelling of oligonucleotides with fluoresceine during automated synthesis. Nucleic Acids Res. 1990;18(11):3427.
[19] Ou CY, Kwok S, Mitchell SW, Mack DH, Sninsky JJ, Krebs JW, Feorino P, Warfield D, Schochetman G. DNA amplification for direct detection of HIV-1 in DNA of peripheral blood mononuclear cells. Science. 1988;239(4837):295-7.