As one of the nucleic acid probes, RNA probes are a class of stretches of single-stranded RNA, which can serve as popular hybridization tools for detecting the presence of complementary target sequences. Normally, RNA probes achieve their detection by being labelled with different tags, such as biotin, fluorophores, etc.
Compared with the most extensively used DNA probes, RNA probes have their own advantages: (a) they can be produced by in vitro transcription, which is a quite economical method to generate large-scale labeled probes with uniform length; (b) they can become the alternative way in nearly all the applications of DNA probes; (c) Some DNA templates cloned in expression vectors can also produce RNA probes, like high specific activity RNA probes or riboprobes; (d) Different with double-stranded DNA probes, single-stranded RNA probes exhibit some merits on signal or hybridization blots. But due to the instability and susceptibility of RNA, they should be treated with the method as other RNA preparations.
RNA probes have many applications in biochemical fields, such as Northern blotting, Southern blotting, in situ hybridization and RNase protection assays. Northern or Southern blotting works by incubating the labeled probes with target DNA or RNA compounds, respectively, followed by visualization of target sequences. For the in situ hybridization experiments, the hybridization and detection of the labeled probes are achieved by localizing of RNA or DNA targets in cells and tissues. In terms of the RNase protection assays, the target RNA can be quantified by the controlled RNase digestion of the hybrids.
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