Abstract
AbstractNonisotopic in situ hybridization can be used to determine the cellular location and the relative levels of expression of specific transcripts within cells and tissues. RNA in prepared specimens is hybridized with a probe labeled nonisotopically with biotin or digoxigenin, which is generally detected by fluorescence or enzymatic methods. Fluorescence in situ hybridization (FISH), probably the most widely used method, is described in addition to techniques for amplification of weak fluorescent signals obtained in FISH. Nonisotopic probes can also be detected by enzymatic reactions using horseradish peroxidase or alkaline phosphatase, as described here.
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