Gel purification of radiolabeled nucleic acids via phosphorimaging: Dip-N-Dot
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 o...
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| Published in: | Analytical biochemistry Vol. 388; no. 2; pp. 351 - 352 |
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| Main Authors: | , , , , , , , , |
| Format: | Journal Article |
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15-05-2009
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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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| AbstractList | RNA and DNA oligonucleotides radiolabeled with (32)P or (33)P 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. RNA and DNA oligonucleotides radiolabeled with [super]32P or [super]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.. 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. |
| Author | Hayden, Eric J. Lehman, Niles Vaidya, Nilesh Arenas, Carolina Díaz Madix, Rowan A. Burton, Aaron S. Chepetan, Andre Larson, Brian C. Riley, Craig A. |
| Author_xml | – sequence: 1 givenname: Aaron S. surname: Burton fullname: Burton, Aaron S. organization: Department of Chemistry, Portland State University, P.O. Box 751, Portland, OR 97207, USA – sequence: 2 givenname: Rowan A. surname: Madix fullname: Madix, Rowan A. organization: Department of Chemistry, Portland State University, P.O. Box 751, Portland, OR 97207, USA – sequence: 3 givenname: Nilesh surname: Vaidya fullname: Vaidya, Nilesh organization: Department of Chemistry, Portland State University, P.O. Box 751, Portland, OR 97207, USA – sequence: 4 givenname: Craig A. surname: Riley fullname: Riley, Craig A. organization: Department of Medicine, Oregon Health and Sciences University, Portland, OR 97239, USA – sequence: 5 givenname: Eric J. surname: Hayden fullname: Hayden, Eric J. organization: Institute of Biochemistry, University of Zürich, CH-8057 Zürich, Switzerland – sequence: 6 givenname: Andre surname: Chepetan fullname: Chepetan, Andre organization: Department of Chemistry, Portland State University, P.O. Box 751, Portland, OR 97207, USA – sequence: 7 givenname: Carolina Díaz surname: Arenas fullname: Arenas, Carolina Díaz organization: Department of Chemistry, Portland State University, P.O. Box 751, Portland, OR 97207, USA – sequence: 8 givenname: Brian C. surname: Larson fullname: Larson, Brian C. organization: Department of Chemistry, Portland State University, P.O. Box 751, Portland, OR 97207, USA – sequence: 9 givenname: Niles surname: Lehman fullname: Lehman, Niles email: niles@pdx.edu organization: Department of Chemistry, Portland State University, P.O. Box 751, Portland, OR 97207, USA |
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| References | Applied Biosystems, Gel purification of probes for nuclease protection assays, Ambion Technical Bulletin no. 171 (2009). Epicentre Biotechnologies, A simple method for RNA purification from in vitro transcription reactions, EPICENTRE Forum 3 (3) (1996). 10.1016/j.ab.2009.02.010_bib1 10.1016/j.ab.2009.02.010_bib2 |
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| Snippet | RNA and DNA oligonucleotides radiolabeled with
32P or
33P often require gel electrophoresis to remove undesired side and/or degradation products. Common ways... RNA and DNA oligonucleotides radiolabeled with (32)P or (33)P often require gel electrophoresis to remove undesired side and/or degradation products. Common... RNA and DNA oligonucleotides radiolabeled with [super]32P or [super]33P often require gel electrophoresis to remove undesired side and/or degradation products.... |
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| SubjectTerms | Autoradiography - methods Electrophoresis - methods Nucleic Acids - analysis Nucleic Acids - chemistry Phosphorus Radioisotopes - chemistry |
| Title | Gel purification of radiolabeled nucleic acids via phosphorimaging: Dip-N-Dot |
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