The determination of DNA based on light-scattering of a complex formed with histone.
The interaction of histone with nucleic acids was characterized by light-scattering measurement using a common spectrofluorometer. Thereby, a sensitive and convenient method for the determination of nucleic acids was established. At pH 4.5-6.5, the interaction of histone with nucleic acids resulted in considerable light-scattering , and four characteristic peaks at 298, 450, 503, and 551 nm were observed. The light-scattering was applied to the determination of nucleic acids. The experiments indicated that, under optimal conditions, a linear relationship was obtained between the light-scattering intensity (I(LS)) and the concentration of nucleic acids. The linear ranges were 0.02-2.0 mug ml(-1) for fish sperm DNA (fsDNA), 0.05-1.5 mug ml(-1) for calf thymus DNA (ctDNA), 0.05-2.5 mug ml(-1) for Herring testis DNA (HtDNA), and 0.05-1.5 mug ml(-1) for human placenta DNA (hpDNA). The detection limits were 2.0 ng for fish sperm DNA, 2.0 ng for calf thymus DNA, 5.0 ng for Herring testis DNA, and 3.0 ng for human placenta DNA. The nucleic acids in yeast cell extraction were determined by simple vortex extraction. The results were satisfactory, and the recovery rates were in the range of 88-108%.
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Research Center for Advanced Science and Technology, The University of Tokyo, 4-6-1 Komaba, Meguro-ku, Tokyo 153-8904, Japan.
Sasaki
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Nakamura
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Karube
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