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RNA Probes: RNA, usually prepared by transcription from cloned DNA, which complements a specific mRNA or DNA and is generally used for studies of virus genes, distribution of specific RNA in tissues and cells, integration of viral DNA into genomes, transcription, etc. Whereas Dna probes are preferred for use at a more macroscopic level for detection of the presence of DNA/RNA from specific species or subspecies, RNA probes are preferred for genetic studies. Conventional labels for the RNA probe include radioisotope labels 32P and 125I and the chemical label biotin. RNA probes may be further divided by category into plus-sense RNA probes, minus-sense RNA probes, and antisense RNA probes.

Understanding Early Organogenesis Using a Simplified In Situ Hybridization Protocol in Xenopus

1Developmental and Stem Cell Biology, Hospital for Sick Children, 2Children's Health Research Institute, University of Western Ontario, 3Department of Physiology and Pharmacology, University of Western Ontario, 4Neurosciences and Mental Health, Hospital for Sick Children, 5Department of Paediatrics, University of Western Ontario

JoVE 51526


 Developmental Biology

Analysis of Single-cell Gene Transcription by RNA Fluorescent In Situ Hybridization (FISH)

1Centre for Medical Parasitology, Department of International Health, Immunology & Microbiology, Faculty of Health Sciences, University of Copenhagen, 2Department of Infectious Diseases, Copenhagen University Hospital (Rigshospitalet), 3Institute of Infection and Immunology Research, School of Biology, University of Edinburgh

JoVE 4073


 Biology

Whole-Mount In Situ Hybridization

JoVE 5330

Whole-mount in situ hybridization (WMISH) is a common technique used for visualizing the location of expressed RNAs in embryos. In this process, synthetically produced RNA probes are first complementarily bound, or "hybridized," to the transcripts of target genes. Immunohistochemistry or fluorescence is then used to detect these RNA hybrids, revealing spatial and temporal patterns of gene expression. Unlike traditional in situ hybridization techniques, which require thin tissue sections whose images will need to be computationally reassembled, the whole-mount technique allows gene expression patterns to be assessed over the entire embryo or structure. This video will introduce the basic concepts of whole mount staining and detail key procedural steps, including probe design and production, embryo fixation and staining, and post-hybridization signal detection. Viewers will then learn about how developmental biologists are applying WMISH to current research studies.


 Developmental Biology

An Introduction to Molecular Developmental Biology

JoVE 5328

Molecular signals play a major role in the complex processes occurring during embryonic development. These signals regulate activities such as cell differentiation and migration, which contribute to the formation of specific cell types and structures. The use of molecular approaches allows researchers to investigate these physical and chemical mechanisms in detail.

This video will review a brief history of the study of molecular events during development. Next, key questions asked by molecular developmental biologists today will be reviewed, followed by a discussion of several prominent methods used to answer these questions, such as staining, explant culture, and live-cell imaging. Finally, we will look at some current applications of these techniques to the study of developmental biology.


 Developmental Biology

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