Abstract
In situ hybridization (ISH) is a widely used technique that has great power in many applications, including diagnosis of viral infections (1), chromosome analysis (2), and mRNA analysis (3). Traditionally, researchers have used radioactive labels to prepare probes for ISH (4). Such probes can be labeled DNA, RNA, or oligonucleotides. RNA and oligonucleotide probes are most often used because they have several features of advantage for ISH (see Table 1); in particular, they are single stranded, offering high sensitivity (RNA) and high-specificity (oligonucleotide). Recently, nonradioactive probes have become more popular, and the same features hold true for these. In this chapter, the ISH process is illustrated by use of a system for the detection of mRNA on tissue sections with RNA probes, but it is equally applicable to work on cell lines and for DNA detection in such systems. In addition, the detection procedures described with the in situ process are applicable to RNA, DNA, and oligonucleotide probes. The analysis of chromosomes and nuclei is a distinct procedure that utilizes nonradioactive DNA probes and fluorescent detection (5).
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© 1996 Humana Press Inc., Totowa, NJ
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Durrant, I. (1996). Nonradioactive In Situ Hybridization for Cells and Tissues. In: Harwood, A.J. (eds) Basic DNA and RNA Protocols. Methods in Molecular Biology™, vol 58. Humana Press. https://doi.org/10.1385/0-89603-402-X:155
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DOI: https://doi.org/10.1385/0-89603-402-X:155
Publisher Name: Humana Press
Print ISBN: 978-0-89603-402-0
Online ISBN: 978-1-59259-251-7
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