Shittu Mujeeb, Steenwinkel Tessa, Dion William, Ostlund Nathan, Raja Komal, Werner Thomas
Department of Biological Sciences, Michigan Technological University, 740 Dow Building, Houghton, MI 49931, USA.
Integrative Systems Biology Graduate Program, University of Pittsburgh School of Medicine, Biomedical Science, Tower 3, 3501 Fifth Avenue, Pittsburgh, PA 15213, USA.
Methods Protoc. 2021 Mar 10;4(1):20. doi: 10.3390/mps4010020.
RNA hybridization (ISH) is used to visualize spatio-temporal gene expression patterns with broad applications in biology and biomedicine. Here we provide a protocol for mRNA ISH in developing pupal wings and abdomens for model and non-model species. We describe best practices in pupal staging, tissue preparation, probe design and synthesis, imaging of gene expression patterns, and image-editing techniques. This protocol has been successfully used to investigate the roles of genes underlying the evolution of novel color patterns in non-model species.
RNA杂交（原位杂交）用于可视化时空基因表达模式，在生物学和生物医学中有广泛应用。在此，我们提供了一种在模型和非模型物种发育中的蛹翅和腹部进行mRNA原位杂交的方案。我们描述了蛹期分期、组织制备、探针设计与合成、基因表达模式成像以及图像编辑技术方面的最佳实践。该方案已成功用于研究非模型物种中新颜色模式进化背后基因的作用。
