A general method is described for distinguishing RNA hairpins from RNA
duplexes by application of two-dimensional filtered nuclear Overhause
r enhancement spectra on a 1:1 mixture of unlabeled and 99% N-15-label
ed molecules. The method is applied to the RNA dodecamer rGGCGCUUGCGUC
which can form an intramolecular hairpin under low salt conditions an
d a duplex in high salt. This procedure allows unambiguous identificat
ion of RNA hairpins or duplexes under the same conditions that are use
d in the NMR solution structure determination.