To confirm that the non-canonical peaks in these 2D experiments were the result of the 2×2 GG/GG internal loop and not a product of stem-stem interactions, we also performed a dilution experiment, where the RNA sample was diluted in buffer and refolded followed by acquiring the 1D spectrum (Figure ). In these experiments, there were no detectable changes upon dilution from 600 µM down to 50 µM, suggesting that these peaks arose from nucleotides in the 2×2 GG/GG internal loop and not from RNA-RNA interactions.