Protocol

In Situ Hybridization Protocols

Volume 326 of the series Methods in Molecular Biology™ pp 89-102

Nonradioactive In Situ Hybridization on Frozen Sections and Whole Mounts

  • Julia Asp
  • , Alexandra Abramsson
  • , Christer Betsholtz

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Summary

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.

Key Words

In situ hybridization gene expression DIG, nonradioactive frozen sections whole mounts mouse embryo