Gnügge Robert, Oh Julyun, Symington Lorraine S
Columbia University Medical Center, New York, NY, United States.
Columbia University Medical Center, New York, NY, United States.
Methods Enzymol. 2018;600:1-24. doi: 10.1016/bs.mie.2017.11.007. Epub 2018 Feb 1.
Single-stranded DNA (ssDNA) intermediates are essential for homology-dependent repair of DNA double-strand breaks (DSBs) and for the DNA damage response. Here we describe methods routinely used to identify ssDNA intermediates formed by end processing of site-specific DSBs in Saccharomyces cerevisiae. These methods have been applied in other model systems and human cell lines, and are useful tools to gain insight into the enzymes that process DSBs and how they are regulated.
单链DNA(ssDNA)中间体对于DNA双链断裂(DSB)的同源依赖性修复和DNA损伤反应至关重要。在这里,我们描述了常用于鉴定酿酒酵母中位点特异性DSB末端加工形成的ssDNA中间体的方法。这些方法已应用于其他模型系统和人类细胞系,是深入了解加工DSB的酶及其调控方式的有用工具。