Analysis of Meiotic Recombination Intermediates by Two-Dimensional Gel Electrophoresis
Analysis of Meiotic Recombination Intermediates by Two-Dimensional Gel Electrophoresis
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DOI：
10.1007/978-1-61779-129-1_7
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发表时间：
2011-01-01
期刊：
影响因子：
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通讯作者：
Boerner, G. Valentin
中科院分区：
文献类型：
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作者：
Ahuja, Jasvinder S.;Boerner, G. Valentin
在减数分裂过程中，程序化的双链断裂产生交叉和非交叉重组产物。减数分裂重组产物是通过几个分支中间体形成的，包括单端侵入和双霍利迪连接。二维凝胶电泳提供了一种灵敏且特异的方法，用于检测分支重组中间体、确定其遗传要求以及富集中间体以供进一步分析。在这里，我们描述了通过二维凝胶电泳对酿酒酵母中的支链重组中间体进行的分析。我们还介绍了减数分裂时间过程、通过链间交联稳定分支 DNA 分子、从减数分裂培养物中提取基因组 DNA 以及二维凝胶印迹的定量分析。
During meiosis, programmed double strand breaks give rise to crossover and non-crossover recombination products. Meiotic recombination products are formed via several branched intermediates, including single end invasions and double Holliday junctions. Two-dimensional gel electrophoresis provides a sensitive and specific approach for detecting branched recombination intermediates, determining their genetic requirements, and enriching intermediates for further analysis. Here, we describe analysis of branched recombination intermediates in the yeast Saccharomyces cerevisiae by two-dimensional gel electrophoresis. We also provide an introduction to meiotic time-course procedures, stabilization of branched DNA molecules by interstrand crosslinking, extraction of gcnomic DNA from meiotic cultures, and quantitative analysis of two-dimensional gel blots.