1. Mung Bean nuclease mapping of RNAs 3' end
- Author
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Giusi Irma Forte, Giorgio L Papadopoulos, Daniele Bellavia, Giorgia Sisino, Rainer Barbieri, Bellavia, D, Sisino, G, Papadopoulos, GL, Forte, GI, and Barbieri, R
- Subjects
lcsh:Immunologic diseases. Allergy ,Aging ,Nuclease ,Mung Bean nuclease ,Hybridization probe ,Immunology ,Short Report ,RNA ,Nuclease protection assay ,Biology ,lcsh:Geriatrics ,Bioinformatics ,Non-coding RNA ,Ageing ,chemistry.chemical_compound ,lcsh:RC952-954.6 ,Biochemistry ,chemistry ,electrophoresis ,Mung Bean Nuclease ,biology.protein ,Small nucleolar RNA ,lcsh:RC581-607 ,DNA - Abstract
A method is described that allows an accurate mapping of 3' ends of RNAs. In this method a labeled DNA probe, containing the presumed 3' end of the RNA under analysis is allowed to anneals to the RNA itself. Mung-bean nuclease is then used to digest single strands of both RNA and DNA. Electrophoretic fractionation of "protected" undigested, labeled DNA is than performed using a sequence reaction of a known DNA as length marker. This procedure was applied to the analysis of both a polyA RNA (Interleukin 10 mRNA) and non polyA RNAs (sea urchin 18S and 26S rRNAs). This method might be potentially relevant for the evaluation of the role of posttrascriptional control of IL-10 in the pathogenesis of the immune and inflammatory mediated diseases associated to ageing. This might allow to develop new strategies to approach to the diagnosis and therapy of age related diseases.
- Published
- 2009