从琼脂糖中制备和分析纯化DNA。
Preparative and analytical purification of DNA from agarose.
作者信息
Vogelstein B, Gillespie D
出版信息
Proc Natl Acad Sci U S A. 1979 Feb;76(2):615-9. doi: 10.1073/pnas.76.2.615.
Abstract
Two procedures were developed for removing DNA from agarose after electrophoretic separation of DNA fragments according to size. Both involve dissolving the DNA-containing agarose in NaI. The preparative technique uses binding of DNA to glass in the presence of NaI. The method is rapid and convenient, and DNA of all molecular weight ranges can be recovered in high yield and without degradation. The DNA is free of agarose and remains susceptible to digestion by restriction enzymes. The analytical technique uses selective precipitation of DNA with acetone and has been adapted to molecular hybridization scans of sequences in agarose gels. The sequence-monitoring system is quantitative, directly measuring the proportion of the probe complementary to a given DNA fragment and vice versa. It is especially suitable for analyzing restriction enzyme digests of DNA in mapping experiments.
摘要
根据大小对DNA片段进行电泳分离后，开发了两种从琼脂糖中去除DNA的方法。两种方法都涉及将含DNA的琼脂糖溶解在碘化钠中。制备技术利用在碘化钠存在下DNA与玻璃的结合。该方法快速简便，所有分子量范围的DNA都能以高产率回收且不发生降解。回收的DNA不含琼脂糖，并且仍可被限制性内切酶消化。分析技术利用丙酮对DNA进行选择性沉淀，并且已适用于琼脂糖凝胶中序列的分子杂交扫描。序列监测系统是定量的，可直接测量与给定DNA片段互补的探针比例，反之亦然。它特别适用于在图谱实验中分析DNA的限制性酶切消化产物。
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