In Situ Hybridization for RNA: Radioactive RNA Probe — A Protocol for Practice With a Kit

Abstract

Recently, nonradioactive probe has often been used to detect RNAs in sectioned tissues by in situ hybridization (ISH) method. Although color-based detection systems with nonradioactive probe is becoming popular in ISH methods, the method with radioactive (RI) probes is still the most sensitive way to detect RNAs in the tissues. Thus, if a given mRNA cannot be detected in tissues with color-based detection methods, although it is confirmed to be present by PCR, the final way to detect the mRNA is to use the ISH method with an RI probe. Processes of specimen preparation, pre-treatments, and hybridization are similar in either radioactive or nonradioactive methods. A major difference between them is the detection method, i.e., autoradiography is used for detection of an RI probe.