Double-label in situ hybridization using biotin and digoxigenin-tagged RNA probes.
Double-label in situ hybridization using biotin and digoxigenin-tagged RNA probes.
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DOI：
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发表时间：
1994-11
期刊：
影响因子：
2.7
通讯作者：
O'Neill Jw;Ethan Bier
中科院分区：
文献类型：
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作者：
O'Neill Jw;Ethan Bier
我们描述了一种基于优化合成生物素标记RNA探针的双标记原位杂交方案。生物素标记的探针与地高辛标记的探针一起使用，同时可视化两种不同的转录本。一个转录物与生物素标记的RNA探针杂交，显示为棕色过氧化物酶反应产物，另一个转录物与地高辛标记的RNA探针杂交，显示为蓝色碱性磷酸酶反应产物。我们提出了几个例子，证明这种双标记方法在确定果蝇胚胎发生过程中表达的各种转录物之间的空间和时间关系方面是有用的，表明这种方法应该普遍用于建立两种独立转录模式的关系。
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.