The widespread availability of recombineered vectors and gene targeted embryonic stem cells from large-scale repositories facilitates the generation of mouse models for functional genetic studies. Southern blotting validates the structure of these targeted alleles produced by homologous recombination, as well as indicating any additional integrations of the vector into the genome. Traditionally this technique employs radioactively-labelled probes; however, there are many laboratories that are restricted in their use of radioactivity. Here, we present a widely applicable protocol for Southern blot analysis using cold probes and alternative procedures employing radioactive probes. Furthermore, the probes are designed to recognise standardised regions of gene-targeting cassettes and so represent universally applicable reagents for assessing allelic integrity.

中文翻译：

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使用冷或放射性探针的通用 Southern 印迹实验方案，用于验证同源重组获得的等位基因


来自大型储存库的重组载体和基因靶向胚胎干细胞的广泛使用促进了用于功能遗传学研究的小鼠模型的生成。 Southern印迹验证了同源重组产生的这些目标等位基因的结构，并表明载体与基因组的任何额外整合。传统上，该技术采用放射性标记的探针。然而，许多实验室在使用放射性方面受到限制。在这里，我们提出了一种广泛适用的使用冷探针和使用放射性探针的替代程序进行 Southern 印迹分析的方案。此外，探针被设计用于识别基因靶向盒的标准化区域，因此代表了用于评估等位基因完整性的普遍适用的试剂。