@article{4156,
  author = {Yu Liu and Erik Holmstrom and Ping Yu and Kemin Tan and Xiaobing Zuo and David Nesbitt and Rui Sousa and Jason Stagno and Yun-Xing Wang},
  title = {Incorporation of isotopic, fluorescent, and heavy-atom-modified nucleotides into RNAs by position-selective labeling of RNA},
  abstract = {Site-specific incorporation of labeled nucleotides is an extremely useful synthetic tool for many structural studies (e.g., NMR, electron paramagnetic resonance (EPR), fluorescence resonance energy transfer (FRET), and X-ray crystallography) of RNA. However, specific-position-labeled RNAs >60 nt are not commercially available on a milligram scale. Position-selective labeling of RNA (PLOR) has been applied to prepare large RNAs labeled at desired positions, and all the required reagents are commercially available. Here, we present a step-by-step protocol for the solid–liquid hybrid phase method PLOR to synthesize 71-nt RNA samples with three different modification applications, containing (i) a 13C15N-labeled segment; (ii) discrete residues modified with Cy3, Cy5, or biotin; or (iii) two iodo-U residues. The flexible procedure enables a wide range of downstream biophysical analyses using precisely localized functionalized nucleotides. All three RNAs were obtained in <2 d, excluding time for preparing reagents and optimizing experimental conditions. With optimization, the protocol can be applied to other RNAs with various labeling schemes, such as ligation of segmentally labeled fragments.},
  year = {2018},
  journal = {Nature Protocols},
  month = {2018-04},
  issn = {1754-2189},
  url = {https://www.nature.com/articles/nprot.2018.002$\#$abstract},
  doi = {10.1038/nprot.2018.002},
}