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. 1997 Dec 1;25(23):4850–4854. doi: 10.1093/nar/25.23.4850

Synthesis and two-dimensional electrophoretic analysis of mixed populations of circular and linear RNAs.

P A Feldstein 1, L Levy 1, J W Randles 1, R A Owens 1
PMCID: PMC147115  PMID: 9365267

Abstract

Spontaneous cleavage of the less abundant form of tobacco ringspot virus satellite RNA is readily reversible. Capitalizing on earlier observations by Feldstein and Bruening that small 'mini-monomer' RNAs derived from this molecule and containing little more than covalently attached ribozyme and substrate cleavage products are able to efficiently circularize, we have constructed a series of self-circularizing RNAs of precisely known size. Mixtures of linear and circular RNAs synthesized in vitro and containing 225-1132 nt could be completely resolved using a novel two-dimensional denaturing polyacrylamide gel electrophoresis system. Similar analyses of a complex mixture of coconut cadang-cadang viroid RNAs revealed the presence of relatively large amounts of a previously undescribed 'fast-slow' heterodimeric RNA species in infected palms. Only a single DNA template is required to prepare each pair of circular and linear RNA markers.

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Selected References

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