scholarly journals Methodology for Whole Mount and Fluorescent RNA In Situ Hybridization in Echinoderms: Single, Double, and Beyond

2020 ◽  
pp. 195-216
Author(s):  
Margherita Perillo ◽  
Periklis Paganos ◽  
Maxwell Spurrell ◽  
Maria I. Arnone ◽  
Gary M. Wessel
Keyword(s):  
In Situ Hybridization ◽  
Whole Mount ◽  
Rna In Situ Hybridization

scholarly journals Whole-Mount RNA In Situ Hybridization and Immunofluorescence of Xenopus Embryos and Tadpoles

2021 ◽  
Vol 2021 (10) ◽  
pp. pdb.prot105635
Author(s):  
Helen Rankin Willsey
Keyword(s):  
In Situ Hybridization ◽  
Final Section ◽  
Stages Of Development ◽  
Xenopus Embryos ◽  
Whole Mount ◽  
Rna In Situ Hybridization ◽  
Endogenous Proteins

A major advantage of experimentation in Xenopus is the ability to query the localization of endogenous proteins and RNAs in situ in the entire animal during all of development. Here I describe three variations of staining to visualize mRNAs and proteins in developing Xenopus embryos and tadpoles. The first section outlines a traditional colorimetric staining for mRNAs that is suitable for all stages of development, and the second extends this protocol for fluorescence-based detection for higher spatial and quantitative resolution. The final section details detection of proteins by immunofluorescence, optimized for tadpole stages but widely applicable to others. Finally, optimization strategies are provided.

Analysis of 3D gene expression patterns in plants using whole-mount RNA in situ hybridization

2014 ◽  
Vol 9 (10) ◽  
pp. 2464-2475 ◽  
Author(s):  
Frédérique Rozier ◽  
Vincent Mirabet ◽  
Teva Vernoux ◽  
Pradeep Das
Keyword(s):  
Gene Expression ◽  
In Situ Hybridization ◽  
Expression Patterns ◽  
Gene Expression Patterns ◽  
Whole Mount ◽  
Rna In Situ Hybridization ◽  
3D Gene

scholarly journals RNA In Situ Hybridization for Detecting Gene Expression Patterns in the Abdomens and Wings of Drosophila Species

2021 ◽  
Vol 4 (1) ◽  
pp. 20
Author(s):  
Mujeeb Shittu ◽  
Tessa Steenwinkel ◽  
William Dion ◽  
Nathan Ostlund ◽  
Komal Raja ◽  
...  
Keyword(s):  
Gene Expression ◽  
In Situ Hybridization ◽  
Expression Patterns ◽  
Drosophila Species ◽  
Gene Expression Patterns ◽  
Probe Design ◽  
Tissue Preparation ◽  
Design And Synthesis ◽  
Rna In Situ Hybridization

RNA in situ 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 Drosophila 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 Drosophila species.

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