Bellavia D et al. (2009), Mung bean nuclease mapping of RNAs 3' end.

ECB-ART-41156

Immun Ageing 2009 May 21;6:6. doi: 10.1186/1742-4933-6-6.

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Mung bean nuclease mapping of RNAs 3'' end.

Bellavia D , Sisino G , Papadopoulos GL , Forte GI , Barbieri R .

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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.

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Genes referenced: LOC100887844

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References [+] :

Cantone, Sequence analysis of the rDNA spacer of Paracentrotus lividus and observations about pre-rRNA processing. NTS sequence of Paracentrotus lividus rDNA. 1993, Pubmed, Echinobase

	Figure 1. Schematic description of Mung Bean mapping method.
	Figure 2. Electrophoretic fractionation of the "protected" labelled probes obtained using our procedure in the mapping of human IL10 RNA (lane 2) and P. lividus 18S and 26S rRNA (lanes 4 and 6, respectively) 3' ends, with respect to a known DNA sequence (in the right). In lanes 1, 3 and 5 the corresponding undigested probes are shown.