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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使用冷探针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

来自大规模储存库的重组载体和基因靶向胚胎干细胞的广泛可用性促进了用于功能遗传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靶向等位基因的结构，并表明载体是否有任何额外的整合到基因组中。传统上，这种技术使用放射性标记的探针；然而，有许多实验室在使用放射性方面受到限制。在这里，我们提出了使用冷探针和使用放射性探针的替代程序进行南方印迹分析的广泛适用的协议。此外，