Kroczek R A
Robert-Koch-Institute, Berlin, Germany.
J Chromatogr. 1993 Aug 25;618(1-2):133-45. doi: 10.1016/0378-4347(93)80031-x.
This review on Southern and Northern analysis, rather than providing step-by-step protocols, focuses on a critical evaluation of the existing experimental methods and modifications thereof. Principal parameters influencing electrophoresis of DNA/RNA in agarose gels are outlined and at the same time alterations in these parameters for optimal resolution of DNA of varying length are discussed. Further, methods for evaluating the quality of DNA/RNA size separation in agarose gels are described. Since efficient transfer of DNA/RNA from the gel onto a membrane support is critical in both methods, several experimental approaches for transfer are compared. Also discussed in this review are alternative methods for radioactive and non-radioactive labeling of DNA probes. Finally, detailed protocols are provided for an effective hybridization of Southern and Northern blots.
本关于Southern和Northern印迹分析的综述，并非提供逐步的实验方案，而是着重对现有实验方法及其改进进行批判性评估。概述了影响DNA/RNA在琼脂糖凝胶中电泳的主要参数，同时讨论了为实现不同长度DNA的最佳分辨率而对这些参数进行的改变。此外，还描述了评估琼脂糖凝胶中DNA/RNA大小分离质量的方法。由于在这两种方法中，DNA/RNA从凝胶有效转移到膜支持物上至关重要，因此比较了几种转移的实验方法。本综述还讨论了DNA探针放射性和非放射性标记的替代方法。最后，提供了Southern和Northern印迹有效杂交的详细实验方案。
