Use of the Ribonuclease Protection Assay for the Analysis and Characterization of Target mRNAs
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Some of the most widely used techniques in the area of molecular biology involve the isolation, analysis, and quantification
of RNA molecules, specifically mRNA molecules that code for proteins of interest. Indeed, the characterization of any gene
entails the analysis of the spatial and temporal distribution of RNA expression. In many types of studies, it is also necessary
to quantify alterations in the synthesis of specific mRNA species that occur both under normal physiological conditions and
in the pathophysiology of diseases such as hypertension. To date, the three most popular methods to characterize RNA molecules
and determine the abundance of a particular mRNA in a total or poly (A) sample are Northern-blot analysis, ribonuclease protection
assays (RPAs), and reverse transcription-polymerase chain reaction (RT-PCR). In theory, each of these techniques can be used
to quantify either the relative or absolute level of an individual RNA species in a population. However, in practice, each
method has inherent technical and practical limitations that may pose significant problems under certain circumstances (1,2)
Affiliation(s): (2) Department of Medicine, College of Human Medicine, Michigan State University, East Lansing, MI
Book Title: Angiotensin Protocols
Series: Methods in Molecular Medicine | Volume: 51 | Pub. Date: Oct-29-2000 | Page Range: 215-222 | DOI: 10.1385/1-59259-087-X:215
Subject: Molecular Medicine
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