S1 nuclease analysis of alternatively spliced mRNA

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  • Molekylærbiologisk Institut
  • Interdisciplinary Nanoscience Center
The characterization of alternatively spliced RNA is a frequently performed task in the molecular biology laboratory. Several methods have been established to characterize specific transcripts, of which microarrays, northern analysis, RT-PCR and nuclease protection assays are the most frequently performed methods in the laboratory. Here, we describe the analysis of alternatively spliced RNA by using 5(')-end labelled DNA oligonucleotide probes and S1 nuclease. The method is sensitive, allowing detection of as little as a few hundred femtograms of a specific RNA, and useful for the quantitation of alternatively spliced mRNA isoforms. Because of its insensitivity towards RNA secondary structures and partially degraded RNA, it may perform better in the quantitation of RNA than northern analysis or RT-PCR, especially when long transcripts are studied.
OriginalsprogEngelsk
TidsskriftMethods in Molecular Biology
Vol/bind703
Sider (fra-til)161-71
Antal sider11
ISSN1064-3745
DOI
StatusUdgivet - 1 jan. 2011

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