Selenium modification of nucleic acids: preparation of oligonucleotides with incorporated 2'-SeMe-uridine for crystallographic phasing of nucleic acid structures.
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.
Pallan
Pradeep S
PS
Department of Biochemistry, School of Medicine, Vanderbilt University, Nashville, Tennessee 37232, USA.
Egli
Martin
M
D002851:Chromatography, High Pressure Liquid; D003461:Crystallography / Q000379:methods*; D015394:Molecular Structure; D009696:Nucleic Acids / Q000737:chemistry*; D015345:Oligonucleotide Probes / Q000138:chemical synthesis* / Q000737:chemistry; D012645:Selenomethionine / Q000737:chemistry*; D014529:Uridine / Q000737:chemistry*
D009696:Nucleic Acids; D015345:Oligonucleotide Probes; D012645:Selenomethionine; D014529:Uridine