SpringerProtocols: Abstract: Identification of G-Protein-Coupled Receptor mRNA Expression by Northern Blotting and In Situ Hybridization

Identification of G-Protein-Coupled Receptor mRNA Expression by Northern Blotting and In Situ Hybridization

By: Lucy F. Donaldson²

Abstract

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G-protein-coupled receptor mRNAs are expressed at low levels and therefore present a challenge for the study of their sites and levels of expression. In situ hybridization (ISH) and Northern blotting are powerful methods for the localization of mRNAs and the study of regulation of mRNA expression. ISH combines the power of precise cellular localization with the ability to perform semiquantitative analysis of the mRNA level, whereas Northern blotting has the ability to identify genetic splice variants, or to study multiple RNA molecules sequentially in the same tissue samples.

These protocols give step-by-step instructions for the performance of these techniques, and the analysis of the data that can be obtained using them.

Affiliation(s): (2) Department of Physiology, University of Bristol School of Medical Sciences, Bristol, UK

Book Title: Receptor Signal Transduction Protocols

Series: Methods in Molecular Biology | Volume: 259 | Pub. Date: Mar-01-2004 | Page Range: 99-122 | DOI: 10.1385/1-59259-754-8:099

Subject: Cell Biology

Key Words: Autoradiography - cRNA probe - in situ hybridization - in vitro transcription - microautoradiog-raphy - northern blotting - poly(A)+ RNA selection, riboprobe, RNase-free conditions

References

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