UVB DNA Dosimeters Analyzed by Polymerase Chain Reactions

Authors

  • Hiroko Yoshida,

    1. Claude Pepper Institute for Aging and Therapeutic Research and Department of Biological Sciences, Florida Institute of Technology, Melbourne, FL, USA
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  • James D. Regan

    Corresponding author
    1. Claude Pepper Institute for Aging and Therapeutic Research and Department of Biological Sciences, Florida Institute of Technology, Melbourne, FL, USA
      *Claude Pepper Institute for Aging and Therapeutic Research, Florida Institute of Technology, 150 West University Boulevard, Melbourne, FL 32901, USA. Fax: 407-952-1818.
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*Claude Pepper Institute for Aging and Therapeutic Research, Florida Institute of Technology, 150 West University Boulevard, Melbourne, FL 32901, USA. Fax: 407-952-1818.

Abstract

Abstract— Purified bacteriophage Λ DNA was dried on a UV-tran-sparent polymer film and served as a UVB dosimeter for personal and ecological applications. Bacteriophage Λ. DNA was chosen because it is commercially available and inexpensive, and its entire sequence is known. Each dosimeter contained two sets of DNA sandwiched between UV-transparent polymer films, one exposed to solar radiation (experimental) and another protected from UV radiation by black paper (control). The DNA dosimeter was then analyzed by a polymerase chain reaction (PCR) that amplifies a 500 base pair specific region of Λ DNA. Photoinduced damage in DNA blocks polymerase from synthesizing a new strand; therefore, the amount of amplified product in UV-exposed DNA was reduced from that found in control DNA. The average lesion frequency per 500 base pair per strand at 16 PCR cycles was –1.22, 1.00, 0.70 and 0.50 for 30 ng, 50 ng, 100 ng and 150 ng of dried DNA, respectively, after a total dose of 60 kj m 2 delivered with a solar UVB simulator. Although the average lesion frequency increases linearly with increasing doses for four different amounts of template DNA, the lesion frequency seems to be averaged by the amplified products from the protected Λ DNA molecules below the top few layers. The average daily dose, equivalent to the UV dose applied with the solar UVB simulator, was 10.2±0.4 kj m2 with the 50 ng containing DNA dosimeter in September 1995 in Melbourne, FL. Both 50 ng and 150 ng containing DNA dosimeters produced the same average daily dose within experimental error in January 1996, which was 5.2±0.3 kj m 2 at the same location. The dried Λ DNA dosimeter is compact, robust, safe and transportable, stable over long storage times and provides the total UVB dose integrated over the exposure time.

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