In Situ Hybridization of Mouse Embryo and Tissue Sections with Radiolabeled RNA Probes
This protocol was adapted from “Techniques for Visualizing Gene Products, Cells, Tissues, and Organ Systems,” Chapter 16, in Manipulating the Mouse Embryo, 3rd edition, by Andras Nagy, Marina Gertsenstein, Kristina Vintersten, and Richard Behringer. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, USA, 2003.INTRODUCTION
This protocol describes in situ hybridization of embryo and tissue sections with 35S-labeled, single-stranded, antisense RNA probes (riboprobes). Protocols have also been developed for in situ hybridization to tissue sections using nonradiolabeled RNA probes that can be detected with antibodies coupled to alkaline phosphatase and a chromogenic substrate. Nonradioactive methods have the advantage that the results can be obtained relatively quickly, but the sensitivity is probably lower than with radioactive probes. In addition, care must be taken to optimize the amount of each probe used in the hybridization reaction.
