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Nonradioactive in situ hybridization on frozen sections and whole mounts.

Journal article
Authors Julia Asp
Alexandra Abramsson
Christer Betsholtz
Published in Methods in molecular biology (Clifton, N.J.)
Volume 326
Pages 89-102
ISSN 1064-3745
Publication year 2006
Published at Institute of Biomedicine, Department of Pathology
Pages 89-102
Language en
Links www.ncbi.nlm.nih.gov/entrez/query.f...
Keywords Animals, Digoxigenin, metabolism, Embryo, cytology, Female, Frozen Sections, Gene Expression, Histological Techniques, In Situ Hybridization, methods, Mice, Molecular Probes, chemical synthesis, Nucleic Acid Hybridization, methods, Pregnancy, RNA Probes, chemical synthesis, RNA, Messenger, metabolism
Subject categories Medical and Health Sciences

Abstract

Nonradioactive in situ hybridization offers a unique opportunity to study gene expression on samples with preserved histological information. This method makes it possible to locate not only where in a tissue a particular gene is expressed, but in many cases also in which specific cell type it is active. Here, we describe our current protocols for in situ hybridization on frozen sections or whole mounts of mouse embryos. The protocols included describe synthesis of a digoxigenin-labeled probe, tissue handling, hybridization of the probe to the mRNA expressed in the sample and signal detection.

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