Short report
Mung Bean nuclease mapping of RNAs 3' end
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* Corresponding author: Rainer Barbieri barner@unipa.it
1 Dipartimento di Biologia Cellulare e dello Sviluppo – Università di Palermo – V.le delle Scienze, Edificio 16, 90128 Palermo, Italy
2 Patologia Clinica, Dipartimento di Biopatologia e Metodologie Biomediche – Università di Palermo, Palermo, Italy
Immunity & Ageing 2009, 6:6 doi:10.1186/1742-4933-6-6
Published: 21 May 2009Abstract
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.