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Production of Digoxigenin‐Labeled Riboprobes for In Situ Hybridization Experiments

Production of Digoxigenin‐Labeled Riboprobes for In Situ Hybridization Experiments 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. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Current Protocols in Mouse Biology Wiley

Production of Digoxigenin‐Labeled Riboprobes for In Situ Hybridization Experiments

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References (17)

Publisher
Wiley
Copyright
© 2020 John Wiley & Sons, Inc.
ISSN
2161-2617
eISSN
2161-2617
DOI
10.1002/cpmo.74
Publisher site
See Article on Publisher Site

Abstract

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.

Journal

Current Protocols in Mouse BiologyWiley

Published: Jun 1, 2020

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