Reinhart Marius, Casas-Delucchi Corella S, Cardoso M Cristina
Department of Biology, Technische Universität Darmstadt, Darmstadt, Germany.
Methods Mol Biol. 2013;1042:213-25. doi: 10.1007/978-1-62703-526-2_15.
The ability of cells to copy their DNA allows them to transmit their genetic information to their progeny. In such, this central biological process preserves the instructions that direct the entire development of a cell. Earlier biochemical analysis in vitro and genetic analysis in yeast laid the basis of our understanding of the highly conserved mechanism of DNA replication. Recent advances on labeling and live-cell microscopy permit now the dissection of this fundamental process in vivo within the context of intact cells. In this chapter, we describe in detail how to perform multiple DNA replication labeling and detection allowing high spatial resolution imaging, as well as how to follow DNA replication in living cells allowing high temporal resolution imaging.
细胞复制其DNA的能力使它们能够将遗传信息传递给后代。因此，这一核心生物学过程保留了指导细胞整个发育过程的指令。早期的体外生化分析和酵母中的遗传分析为我们理解高度保守的DNA复制机制奠定了基础。最近在标记和活细胞显微镜技术方面的进展，现在使我们能够在完整细胞的背景下在体内剖析这一基本过程。在本章中，我们将详细描述如何进行多重DNA复制标记和检测，以实现高空间分辨率成像，以及如何在活细胞中追踪DNA复制，以实现高时间分辨率成像。
