Selenium modification of nucleic acids: preparation of oligonucleotides with incorporated 2'-SeMe-uridine for crystallographic phasing of nucleic acid structures

Nat Protoc. 2007;2(3):647-51. doi: 10.1038/nprot.2007.75.

Abstract

This protocol describes a method for introducing an anomalously scattering atom into oligonucleotides at the 2'-position of uridine by conventional solid-phase synthesis. The 2'-SeMe ribose modification is particularly attractive for derivatization of RNA to facilitate crystal structure determination. The estimated time for the synthesis and HPLC purification of oligonucleotides with incorporated 2'-SeMe-uridine residues is approximately 46 h for 'trityl on' and approximately 32 h for 'trityl off' methods, respectively.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Chromatography, High Pressure Liquid
  • Crystallography / methods*
  • Molecular Structure
  • Nucleic Acids / chemistry*
  • Oligonucleotide Probes / chemical synthesis*
  • Oligonucleotide Probes / chemistry
  • Selenomethionine / chemistry*
  • Uridine / chemistry*

Substances

  • Nucleic Acids
  • Oligonucleotide Probes
  • Selenomethionine
  • Uridine