O'Neill J W, Bier E
University of California, San Diego, La Jolla.
Biotechniques. 1994 Nov;17(5):870, 874-5.
We describe a double-label in situ hybridization protocol based on the optimized synthesis of biotin-labeled RNA probes. Biotin-labeled probes are used in conjunction with digoxigenin-labeled probes to simultaneously visualize two different transcripts. One transcript is hybridized with a biotin-labeled RNA probe and visualized as a brown peroxidase reaction product, and the other transcript is hybridized with a digoxigenin-labeled RNA probe and visualized as a blue alkaline phosphatase reaction product. We present several examples in which this double-labeling method has proven useful in determining the spatial and temporal relationships between various transcripts expressed during Drosophila embryogenesis, indicating that this method should be of general use in establishing the relationship of two independent transcription patterns.
我们描述了一种基于生物素标记RNA探针优化合成的双标记原位杂交方案。生物素标记的探针与地高辛配体标记的探针联合使用，以同时可视化两种不同的转录本。一种转录本与生物素标记的RNA探针杂交，并可视化为棕色过氧化物酶反应产物，另一种转录本与地高辛配体标记的RNA探针杂交，并可视化为蓝色碱性磷酸酶反应产物。我们给出了几个例子，其中这种双标记方法已被证明在确定果蝇胚胎发育过程中表达的各种转录本之间的空间和时间关系方面很有用，这表明该方法在建立两种独立转录模式的关系方面应具有普遍用途。
