Paredes Aaron J, Alfaro-Valdés Hilda M, Wilson Christian A M
Departamento de Bioquímica y Biología Molecular, Facultad de Ciencias Químicas y Farmacéuticas, Universidad de Chile.
Departamento de Bioquímica y Biología Molecular, Facultad de Ciencias Químicas y Farmacéuticas, Universidad de Chile;
J Vis Exp. 2018 Jan 28(131):56528. doi: 10.3791/56528.
DNA staining methods are very important for biomedical research. We designed a simple method that allows DNA visualization to the naked eye by the formation of a colored precipitate. It works by soaking the acrylamide or agarose DNA gel in a solution of 1x (equivalent to 2.0 µM) SYBR Green I (SG I) and 0.20 mM nitro blue tetrazolium that produces a purple precipitate of formazan when exposed to sunlight or specifically blue light. Also, DNA recovery tests were performed using an ampicillin resistant plasmid in an agarose gel stained with our method. A larger number of colonies was obtained with our method than with traditional staining using SG I with ultraviolet illumination. The described method is fast, specific, and non-toxic for DNA detection, allowing visualization of biomolecules to the "naked eye" without a transilluminator, and is inexpensive and appropriate for field use. For these reasons, our new DNA staining method has potential benefits to both research and industry.
DNA染色方法对生物医学研究非常重要。我们设计了一种简单的方法,通过形成有色沉淀使DNA肉眼可见。其原理是将丙烯酰胺或琼脂糖DNA凝胶浸泡在含有1x(相当于2.0 µM)的SYBR Green I(SG I)和0.20 mM硝基蓝四唑的溶液中,当暴露在阳光或特定蓝光下时会产生紫色的甲臜沉淀。此外,我们使用氨苄青霉素抗性质粒在经我们方法染色的琼脂糖凝胶中进行了DNA回收测试。与使用SG I和紫外线照明的传统染色方法相比,我们的方法获得了更多的菌落。所描述的方法对DNA检测快速、特异且无毒,无需透射仪即可使生物分子肉眼可见,而且成本低廉,适合现场使用。基于这些原因,我们新的DNA染色方法对研究和工业都有潜在的益处。