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Rapid NMR screening of RNA secondary structure and binding
Ist Teil von
Journal of biomolecular NMR, 2015-09, Vol.63 (1), p.67-76
Ort / Verlag
Dordrecht: Springer Netherlands
Erscheinungsjahr
2015
Link zum Volltext
Quelle
SpringerLink (Online service)
Beschreibungen/Notizen
Determination of RNA secondary structures by NMR spectroscopy is a useful tool e.g. to elucidate RNA folding space or functional aspects of regulatory RNA elements. However, current approaches of RNA synthesis and preparation are usually time-consuming and do not provide analysis with single nucleotide precision when applied for a large number of different RNA sequences. Here, we significantly improve the yield and 3′ end homogeneity of RNA preparation by in vitro transcription. Further, by establishing a native purification procedure with increased throughput, we provide a shortcut to study several RNA constructs simultaneously. We show that this approach yields μmol quantities of RNA with purities comparable to PAGE purification, while avoiding denaturation of the RNA.