Rybak-Wolf Agnieszka, Solana Jordi
Laboratory for Systems Biology of Gene Regulatory Elements, Berlin Institute for Medical Systems Biology, Max-Delbrück-Center for Molecular Medicine, Robert-Rössle-Strasse 10, 13125, Berlin, Germany,
Methods Mol Biol. 2014;1211:41-51. doi: 10.1007/978-1-4939-1459-3_4.
In recent years freshwater flatworms (planarian) have become a powerful model for studies of regeneration and stem cell biology. Whole-mount in situ hybridization (WISH) and fluorescent in situ hybridization (FISH) are key and most commonly used techniques to determine and visualize gene expression patterns in planaria. Here, we present the established version of whole-mount in situ hybridization (WISH) and whole-mount fluorescence in situ hybridization (WFISH) protocol optimized over the last years by several labs from the rapidly growing planaria field and give an overview of recently introduced modifications which can be critical in the study of low abundant transcripts.
近年来,淡水扁虫(涡虫)已成为研究再生和干细胞生物学的强大模型。全组织原位杂交(WISH)和荧光原位杂交(FISH)是确定和可视化涡虫基因表达模式的关键且最常用的技术。在此,我们展示了全组织原位杂交(WISH)和全组织荧光原位杂交(WFISH)方案的成熟版本,该方案是近年来由快速发展的涡虫研究领域的多个实验室优化而来的,并概述了最近引入的一些修改,这些修改在低丰度转录本的研究中可能至关重要。