Gel purification of radiolabeled nucleic acids via phosphorimaging: Dip-N-Dot

• Published in: Analytical biochemistry Vol. 388; no. 2; pp. 351 - 352
• Main Authors: Burton, Aaron S., Madix, Rowan A., Vaidya, Nilesh, Riley, Craig A., Hayden, Eric J., Chepetan, Andre, Arenas, Carolina Díaz, Larson, Brian C., Lehman, Niles
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 15-05-2009
• Subjects: Autoradiography - methods; Electrophoresis - methods; Nucleic Acids - analysis; Nucleic Acids - chemistry; Phosphorus Radioisotopes - chemistry
• ISSN: 0003-2697; 1096-0309
• DOI: 10.1016/j.ab.2009.02.010

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.