Department of Environmental & Radiological Health Sciences, Colorado State University, Fort Collins, CO, USA.
Methods Mol Biol. 2023;2519:111-116. doi: 10.1007/978-1-0716-2433-3_13.
The standard FISH uses DNA probes to hybridize to the designated complementary strands. This is DNA-DNA interaction, and it usually takes much longer time to obtain detectable signals compared to other reactions such as immunochemical reactions and simple chemical reactions. Certain proteins bind to specific DNA sequences and regulate the biological function of DNA. These DNA-binding proteins have specific domains to interact with single- or double-stranded DNA. Some of telomere proteins apparently bind to telomere sequence and form nucleoprotein complex to protect chromosome ends. Using telomere PNA probes, probes can be accumulated at the telomere sites in a non-hybridization manner. This chapter introduces nontraditional PNA telomere staining protocol without DNA-DNA hybridization to visualize telomere locations on metaphase chromosomes.
标准的 FISH 使用 DNA 探针与指定的互补链杂交。这是 DNA-DNA 相互作用,与其他反应(如免疫化学反应和简单的化学反应)相比,通常需要更长的时间才能获得可检测的信号。某些蛋白质与特定的 DNA 序列结合并调节 DNA 的生物学功能。这些 DNA 结合蛋白具有与单链或双链 DNA 相互作用的特定结构域。一些端粒蛋白显然与端粒序列结合形成核蛋白复合物以保护染色体末端。使用端粒 PNA 探针,探针可以以非杂交的方式在端粒部位积累。本章介绍了一种非传统的 PNA 端粒染色方案,无需 DNA-DNA 杂交即可在中期染色体上可视化端粒位置。