Production of Digoxigenin-Labeled Riboprobes for In Situ Hybridization Experiments.
RNA detection
digoxigenin
fluorescein
gene expression
hybridization
mouse embryo
riboprobe
Journal
Current protocols in mouse biology
ISSN: 2161-2617
Titre abrégé: Curr Protoc Mouse Biol
Pays: United States
ID NLM: 101560384
Informations de publication
Date de publication:
Jun 2020
Jun 2020
Historique:
entrez:
22
5
2020
pubmed:
22
5
2020
medline:
26
1
2021
Statut:
ppublish
Résumé
Experiments that visualize gene expression in intact tissues or organisms are fundamental to studies of gene function. These experiments, called in situ hybridization, require the production of a riboprobe, which is a labeled antisense RNA corresponding to a particular gene. The most commonly used system for visualizing gene expression via in situ hybridization is the incorporation of a digoxigenin label into an in vitro-transcribed RNA probe. After hybridization of the riboprobe to a target mRNA, its location can be detected via a high-affinity α-digoxigenin antibody conjugated to an alkaline-phosphatase enzyme. The article describes the design and production of digoxigenin-labeled riboprobes transcribed in vitro from template DNA (either plasmid or PCR amplicon). These riboprobes are suitable for use in tissue and whole-mount in situ hybridization protocols. © 2020 by John Wiley & Sons, Inc. Basic Protocol 1: Plasmid-derived riboprobes Alternate Protocol: PCR-derived riboprobes Basic Protocol 2: Riboprobe synthesis with DIG label.
Substances chimiques
RNA Probes
0
Digoxigenin
NQ1SX9LNAU
Fluorescein
TPY09G7XIR
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
e74Subventions
Organisme : National Health and Medical Research Council
ID : 1126288
Informations de copyright
© 2020 John Wiley & Sons, Inc.
Références
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