Whole Mount in Situ Hybridization for mRNA Detection in Chick Embryos
Analysis of the spatial expression patterns of genes in animal embryos is essential to understanding functions of the genes especially for developmental biology. In situ hybridization with whole embryos is a powerful method to observe overall expression patterns of genes in embryos. A method for the whole mount in situ hybridization was developed after non-radioactive probes, especially digoxigenin-labeled RNA probes, became commercially available. Furthermore, using two different non-radioactive probes, expression patterns of two different genes can be observed simultaneously in an embryo, as a pattern of two colors. Sections of the stained embryos can be used to identify cells expressing the mRNA in the embryo. Thus, this method allows the identification of mRNA within intact embryos at both regional and cellular levels.
KeywordsChick Embryo Spatial Expression Pattern mRNA Detection Nitroblue Tetrazolium Chloride Wash Pellet
Unable to display preview. Download preview PDF.
- Noji S. (1997) A practical protocol for whole-mount in situ hybridization. In: Immunostaining and In situ hybridization (Yodosha The Protocol Series), pp80–93. Tokyo: YodoshaGoogle Scholar
- Wilkinson D.G. (1992). Whole mount in situ hybridization of vertebrate embryos. In: In Situ Hybridization, A Practical Approach (ed.D.G. Wilkinson), pp.75–83. Oxford: IRL press at Oxford University PressGoogle Scholar