Goffart Steffi, Pohjoismäki Jaakko
University of Eastern Finland, Department of Environmental and Biological Sciences, Joensuu, Finland.
Methods Mol Biol. 2023;2615:241-266. doi: 10.1007/978-1-0716-2922-2_18.
Two-dimensional neutral/neutral agarose gel electrophoresis (2D-AGE) has been employed for nearly two decades in the analysis of replication and maintenance processes of animal mitochondrial DNA, but the method's potential has not been fully exploited. Here, we describe the various steps involved in this technique, from DNA isolation, to two-dimensional neutral/neutral agarose gel electrophoresis (2D-AGE), Southern hybridization and interpretation. We also provide examples of the applicability of 2D-AGE to investigate the different features of mtDNA maintenance and regulation.
二维中性/中性琼脂糖凝胶电泳（2D-AGE）已被用于动物线粒体DNA复制和维持过程分析近二十年，但该方法的潜力尚未得到充分发掘。在此，我们描述了这项技术所涉及的各个步骤，从DNA分离到二维中性/中性琼脂糖凝胶电泳（2D-AGE）、Southern杂交及结果解读。我们还提供了二维中性/中性琼脂糖凝胶电泳（2D-AGE）用于研究线粒体DNA维持和调控不同特征的适用性示例。
