Improved mass analysis of oligoribonucleotides by 13C, 15N double depletion and electrospray ionization FT-ICR mass spectrometry.
13C, 15N doubly depleted 32-ribonucleotide was synthesized enzymatically by in vitro transcription from nucleoside triphosphates isolated from E. coli grown in a minimal medium containing 12C, 14N-enriched glucose and ammonium sulfate. Following purification and desalting by reversed-phase HPLC, buffer exchange with Microcon YM-3, and ethanol precipitation, electrospray ionization Fourier transform ion cyclotron resonance mass spectra revealed greatly enhanced abundance of monoisotopic ions (by a factor of approximately 100) and a narrower isotopic distribution with higher signal-to-noise ratio. The abrupt onset and high magnitude of the monoisotopic species promise to facilitate accurate mass measurement of RNA's.
Xiong
Ying
Y
Institute of Molecular Biophysics, Florida State University, Tallahassee, Florida 32306, USA.
Schroeder
Kersten
K
Greenbaum
Nancy L
NL
Hendrickson
Christopher L
CL
Marshall
Alan G
AG
D002247:Carbon Isotopes; D005583:Fourier Analysis; D009587:Nitrogen Isotopes; D009843:Oligoribonucleotides / Q000032:analysis* / Q000737:chemistry*; D012265:Ribonucleotides / Q000032:analysis / Q000737:chemistry; D012492:Salts / Q000737:chemistry; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*; D017550:Spectroscopy, Fourier Transform Infrared / Q000379:methods*
D002247:Carbon Isotopes; D009587:Nitrogen Isotopes; D009843:Oligoribonucleotides; D012265:Ribonucleotides; D012492:Salts