改良琼脂糖凝胶用于短 DNA 片段的高效 Southern 印迹分析。

Modified agarose gel for high-performance Southern blotting of short DNA fragments.

机构信息

Laboratory of Molecular and Developmental Biology, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing, China.

出版信息

Biotechniques. 2010 Aug;49(2):580-1. doi: 10.2144/000113465.

DOI:10.2144/000113465

PMID:20701593

Abstract

Southern blotting is a common method used for the study of gene organization. Current methods of DNA transfer for Southern blotting, however, can be inefficient for high concentration agarose gels. Here, we report a method for high-performance Southern blotting of short DNA fragments such as nucleosomal DNAs by using a discontinuous agarose zone gel. The results show that sharp and well-resolved fractionation of short DNA fragments comparable to that from a high-concentration agarose gel could be obtained using a low-concentration agarose gel with a small zone of high-concentration agarose, and that the resulting DNA transfer is highly efficient and rapid.

摘要

Southern 印迹法是一种常用于研究基因组织的常用方法。然而，目前用于 Southern 印迹法的 DNA 转移方法对于高浓度琼脂糖凝胶可能效率不高。在这里，我们报告了一种通过使用不连续琼脂糖区带凝胶对短 DNA 片段（如核小体 DNA）进行高效 Southern 印迹法的方法。结果表明，使用低浓度琼脂糖凝胶和小区域高浓度琼脂糖，可以获得与高浓度琼脂糖凝胶相当的短 DNA 片段的锐利且分辨率良好的分离，并且所得的 DNA 转移非常高效和快速。