Gel purification of radiolabeled nucleic acids via phosphorimaging: Dip-N-Dot |
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Authors: | Aaron S Burton Rowan A Madix Nilesh Vaidya Craig A Riley Eric J Hayden Andre Chepetan Carolina Díaz Arenas Brian C Larson Niles Lehman |
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Institution: | 1. Department of Chemistry, Portland State University, P.O. Box 751, Portland, OR 97207, USA;2. Department of Medicine, Oregon Health and Sciences University, Portland, OR 97239, USA;3. Institute of Biochemistry, University of Zürich, CH-8057 Zürich, Switzerland |
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Abstract: | RNA and DNA oligonucleotides radiolabeled with 32P or 33P often require gel electrophoresis to remove undesired side and/or degradation products. Common ways to visualize these molecules after electrophoresis are by ultraviolet (UV) shadowing, which necessarily reduces the specific activity of the oligonucleotide, and by autoradiography using film, which is cumbersome and increases the cost of generating the radiolabeled molecule. A more cost-effective method is to physically inject the gel with a “Dip-N-Dot” solution of dye and radionuclide after electrophoresis but prior to phosphorimaging. The gel can be overlaid on its computer-generated image, allowing the labeled molecules to be visualized quickly. |
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