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首页> 外文期刊>Proceedings of the National Academy of Sciences of the United States of America >Trapping of megabase-sized DNA molecules during agarose gel electrophoresis
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Trapping of megabase-sized DNA molecules during agarose gel electrophoresis

机译：琼脂糖凝胶电泳过程中诱捕兆碱基大小的DNA分子

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摘要

Megabase DNA molecules become trapped in agarose gels during electrophoresis if the electric field exceeds a few volts per cm. Fluorescence microscopy reveals that these molecules invariably arrest in U-shaped conformations. The field-vs.-size dependence for trapping indicates that a critical molecular tension is required for trapping. The size of unli- gated #lambda#-ladders, sheared during gel electrophoresis at a given field, coincides with the size of molecules trapped at that field, suggesting that both processes occur through nick melting near the vertex of the U-shape. Consistently, molecules nicked by exposure to UV radiation trap more readily than unexposed ones. The critical trapping tension at the vertex is estimated to be 15 pN, a force sufftcient to melt nicks bent around gel fibers, and, according to our model, trap a molecule. Strate- gies to reduce. molecular tension and avoid trapping are discussed.
机译：如果电场超过每厘米几伏，则在电泳过程中，megabase DNA分子将被捕获在琼脂糖凝胶中。荧光显微镜显示，这些分子总是以U形构型阻滞。捕获的场-尺寸依赖性表明捕获需要临界分子张力。在给定电场的凝胶电泳过程中剪切的未包封的＃lambda＃梯形的大小与在该电场上捕获的分子的大小相吻合，表明这两个过程都是通过在U形顶点附近的切口熔化而发生的。始终如一，与未曝光的分子相比，因暴露于紫外线辐射而形成切口的分子更容易捕获。顶点处的临界捕获张力估计为15 pN，该力足以融化凝胶纤维周围弯曲的缺口，并根据我们的模型捕获分子。减少策略。讨论了分子张力和避免捕获。

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