Application of a Novel Fluorescence Probe in the Determination of Nucleic Acids

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Qing-Zhi Zhu, Fang Li, Xiang-Qun Guo, Jin-Gou Xu and Wen-You Li


Abstract

A novel fluorimetric method has been developed for rapid determination of DNA and RNA with hypocrellin A (HA) as a fluorescence probe, based on the fluorescence enhancement of HA in the presence of DNA or RNA. Maximum fluorescence is produced in the pH range 3.4–4.0, with maximum excitation and emission wavelengths at 470 and 600 nm, respectively. Under optimal conditions, the calibration graphs are linear over the range 0–200.0 ng cm-3 for calf thymus DNA and 13.0–200.0 ng cm-3 for yeast RNA, respectively. The corresponding detection limits are 5.0 ng cm-3 for calf thymus DNA and 13.0 ng cm-3 for yeast RNA. The relative standard deviation of six replicate measurements is 4.5% for 100 ng cm-3 calf thymus DNA. DNA could be determined in the presence of 20% m/m yeast RNA. The mechanism for the binding of HA to DNA is also studied.


References

  1. S. Udenfriend and P. Zaltzman, Anal. Biochem., 1962, 3, 49 CrossRef CAS.
  2. H. C. Borresen, Acta Chem. Scand., 1963, 17, 921 CAS.
  3. J. R. Lakowicz, Fluorescence Spectroscopy, Vol 3: Biochemical Applications, Plenum Press, New York, 1992, pp. 137 Search PubMed.
  4. J.-B. Le Pecq and C. Paoletli, Anal. Biochem., 1966, 17, 100 CrossRef CAS.
  5. J. Kapuscinski and B. Skoczylas, Anal. Biochem., 1977, 83, 252 CrossRef CAS.
  6. C. Labarca and K. Paigen, Anal. Biochem., 1980, 102, 344 CrossRef CAS.
  7. M. D. Topal and J. R. Fresco, Biochemistry, 1980, 19, 5531 CrossRef CAS.
  8. M. Al-Hakeem and S. S. Sommer, Anal. Biochem., 1987, 163, 433 CrossRef CAS.
  9. D. Pavlick and C. Formoso, Biochemistry, 1978, 17, 1537 CrossRef CAS.
  10. D. P. Ringer, B. A. Howell and D. E. Kizer, Anal. Biochem., 1980, 103, 337 CAS.
  11. Y. X. Ci, Y. Z. Li and W. B. Chang, Anal. Chim. Acta, 1991, 248, 589 CrossRef CAS.
  12. Y. X. Ci, Y. Z. Li and X. J. Liu, Anal. Chem., 1995, 67, 1785 CrossRef CAS.
  13. H. S. Rye, S. Yue, D. E. Wemmer, M. A. Quesada, R. P. Haugland, R. A. Mathies and A. N. Glazer, Nucleic Acids Res., 1992, 20, 2803 CrossRef CAS.
  14. H. S. Rye, J. M. Dabora, M. A. Quesada, R. A. Mathies and A. N. Glazer, Anal. Biochem., 1993, 208, 144 CrossRef CAS.
  15. X. Y. Wan and Y. T. Cheng, Chinese Sci. Bull., 1980, 25, 1148 Search PubMed.
  16. L. J. Jiang, Chinese Sci. Bull., 1990, 35, 1608 Search PubMed.
  17. L. J. Jiang, Chinese Sci. Bull., 1990, 35, 1681 Search PubMed.
  18. Z. J. Diwu and J. W. Lown, J. Photochem. Photobiol. A: Chem., 1992, 64, 273 CrossRef CAS.
  19. Z. J. Diwu and J. W. Lown, J. Photochem. Photobiol. A: Chem., 1992, 69, 191 CrossRef CAS.
  20. Z. J. Diwu, C. Zhang and J. W. Lown, J. Photochem. Photobiol. A: Chem., 1992, 66, 99 CrossRef CAS.
  21. Z. J. Diwu and J. W. Lown, Photochem. Photobiol., 1990, 52, 609 Search PubMed.
  22. Z. J. Diwu, L. J. Jiang and M. H. Zhang, Sci. Sin., Ser. B (Engl. Ed.), 1989, 33, 18 Search PubMed.
  23. Z. J. Diwu, L. J. Jiang and M. H. Zhang, Acta Phys. Chem. Sin., 1989, 5, 250 Search PubMed.
  24. J.-B. Le Pecq and C. Paoletti, C. R. Acad. Sci. Paris, 1964, 259, 1786 Search PubMed.
  25. W. D. Wilson, F. A. Tanious, R. A. Watson, H. J. Barton, A. Strekowska, D. B. Harden and L. Strekowski, Biochemistry, 1989, 28, 1984 CrossRef CAS.
  26. C. Cantor and P. R. Schimmel, Biophysical Chemistry, Freeman, San Francisco, 1980, vol. 2, pp. 398 Search PubMed.
  27. C. V. Kumar, R. S. Turner and E. H. Asuncion, J. Photochem. Photobiol. A: Chem., 1993, 74, 231 CrossRef CAS.
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