Optimized Preparation of Segmentally Labeled RNAs for NMR Structure Determination.

RNA structures are significantly underrepresented in public repositories (∼ 100-fold compared to proteins) despite their importance for mechanistic understanding and for development of structure prediction/validation tools. A substantial portion of deposited RNA structures have been determined by NMR (∼ 30%), but most comprise fewer than 60 nucleotides due to complications associated with NMR signal overlap. A promising approach for applying NMR to larger RNAs involves use of a mutated DNA polymerase (TGK) that can extend "primer" RNA strands generated independently by synthetic or enzymatic methods [Haslecker et al.