SEPARATION OF VERY LARGE DNA-MOLECULES BY GEL-ELECTROPHORESIS
SEPARATION OF VERY LARGE DNA-MOLECULES BY GEL-ELECTROPHORESIS
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DOI：
10.1093/nar/5.3.653
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发表时间：
1978-01-01
影响因子：
14.9
通讯作者：
FANGMAN, WL
中科院分区：
文献类型：
--
作者：
FANGMAN, WL
通过在稀琼脂糖水平板凝胶中电泳分离非常大的 DNA 分子。电泳条件是使用来自细菌病毒 λ、T4 和 G 的完整 DNA 分子开发的。它们的 DNA 的分子量 (M) 分别为 3200 万、1.2 亿和 5 亿。已发现多种电泳条件可提供足够高的迁移率和大的迁移率差异，从而使这些 DNA 在短时间内得以分离。在 0.1% 琼脂糖中以 2.5 V/cm 凝胶长度进行电泳，只需 10 小时即可将 T4 和 λ DNA 分离 2.0 cm，将 G 和 T4 DNA 分离 1.0 cm。在某些条件下，对于 M 值从 10 到 5 亿，DNA 迁移率与 log M 成正比。程序。使用该技术可以快速测定分子量并分离非常大的 DNA 分子。
Very large DNA molecules were separated by electrophoresis in horizontal slab gels of dilute agarose. Conditions of electrophoresis were developed using intact DNA molecules from the bacterial viruses λ, T4 and G. Their DNAs have molecular weights (M) of 32 million, 120 million, and 500 million, respectively. Several electrophoresis conditions were found which give sufficiently high mobilities and large mobility differences that these DNAs are separated in a short time. Electrophoresis in 0.1% agarose at 2.5 V/cm of gel length separates T4 and λ DNAs by 2.0 cm, and G and T4 DNAs by 1.0 cm in only 10 hr. With some conditions DNA mobilities are directly proportional to log M for M values from 10 to 500 million. The procedures. used will allow rapid molecular weight determination and separation of very large DNA molecules.