Synthesis of backbone deuterium labelled [r(CGCGAAUUCGCG)]2 and HPLC purification of synthetic RNA.
The chemical synthesis of backbone deuterium labelled [r(CGCGAAU*U*CGCG)]2 (U* = [5'-2H]U) is described. An efficient purification procedure was developed using a polymeric reverse phase (PRP) HPLC column at 60 degrees C. This procedure provided pure RNA dodecamer in the multi-milligram quantities (39% overall yield) necessary for dynamics studies using solid-state deuterium NMR. The purification method has been effectively applied to other RNA sequences and will assist biophysical studies which require relatively large quantities of RNA oligomers.
Khare
D
D
Maryland Biotechnology Institute, University of Maryland, Rockville 20850.
Orban
J
J
D000577:Amides; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D003903:Deuterium; D007202:Indicators and Reagents; D007553:Isotope Labeling / Q000379:methods; D009682:Magnetic Resonance Spectroscopy / Q000379:methods; D009843:Oligoribonucleotides / Q000138:chemical synthesis* / Q000302:isolation & purification; D010756:Phosphoric Acids; D012313:RNA / Q000138:chemical synthesis* / Q000302:isolation & purification
D000577:Amides; D007202:Indicators and Reagents; D009843:Oligoribonucleotides; D010756:Phosphoric Acids; D012313:RNA; C011067:phosphoramidic acid; D003903:Deuterium