Using Pulsed-Field Gel Electrophoresis to Analyze Schizosaccharomyces pombe Chromosomes and Chromosomal Elements.
Using Pulsed-Field Gel Electrophoresis to Analyze Schizosaccharomyces pombe Chromosomes and Chromosomal Elements.
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使用脉冲场凝胶电泳分析粟酒裂殖酵母染色体和染色体元素。
DOI：
10.1101/pdb.prot092023
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发表时间：
2018
影响因子：
--
通讯作者：
Pai CC
中科院分区：
文献类型：
--
作者：
Pai CC
脉冲场凝胶电泳（PFGE）使用交替定向的脉冲电场来分离大的DNA分子。在这里，我们描述了PFGE分离和可视化0.5 - 6mb染色体（最适合分析内源性裂变酵母染色体5.7,4.6和3.5 Mb）的方案和条件，以及50 - 600 kb之间较短的染色体元件，如530 kb的ch16小染色体。除了确定染色体大小外，该技术还具有广泛的应用，包括确定DNA复制或修复是否完成，定义细胞的分子核型，分析染色体重排，将基因或结构分配到特定染色体，以及从特定染色体中分离DNA。
Pulsed field gel electrophoresis (PFGE) uses alternatively oriented pulsed electrical fields to separate large DNA molecules. Here, we describe PFGE protocols and conditions for separating and visualizing chromosomes between 0.5 and 6 Mb (optimal for analyzing the endogenous fission yeast chromosomes of 5.7, 4.6, and 3.5 Mb), and for shorter chromosomal elements of between 50 and 600 kb, such as the 530 kb Ch 16 minichromosome. In addition to determining chromosome size, this technique has a wide range of applications, including determining whether DNA replication or repair is complete, defining the molecular karyotype of cells, analyzing chromosomal rearrangements, assigning genes or constructs to particular chromosomes, and isolating DNA from specific chromosomes.