Matrix assisted laser desorption/ionization mass spectrometry of enzymatically synthesized RNA up to 150 kDa

F Kirpekar; E Nordhoff; K Kristiansen; P Roepstorff; A Lezius; S Hahner; M Karas; F Hillenkamp

doi:10.1093/nar/22.19.3866

Matrix assisted laser desorption/ionization mass spectrometry of enzymatically synthesized RNA up to 150 kDa

Nucleic Acids Res. 1994 Sep 25;22(19):3866-70. doi: 10.1093/nar/22.19.3866.

Authors

F Kirpekar¹, E Nordhoff, K Kristiansen, P Roepstorff, A Lezius, S Hahner, M Karas, F Hillenkamp

Affiliation

¹ Department of Molecular Biology, Odense University, Denmark.

Abstract

Enzymatically synthesized RNA samples (in vitro transcripts) were analysed by matrix assisted laser desorption/ionization mass spectrometry (MALDI-MS). Spectra of RNA up to 150 kDA (461 nucleotides) are shown. Polymerase generated sample heterogeneity and its contribution to mass resolution are discussed. A time course exonuclease digest of a 55 nt in vitro transcript was analyzed to investigate the performance of MALDI-MS on complex mixtures. Based on these data, the analysis by MALDI-MS of DNA sequencing reactions, produced by the action of an RNA polymerase, is discussed.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Base Sequence
DNA-Directed RNA Polymerases / metabolism
Lasers
Mass Spectrometry*
Molecular Sequence Data
Molecular Weight
Phosphoric Diester Hydrolases / metabolism
RNA / analysis*
RNA / biosynthesis
RNA / chemistry
Sequence Analysis, RNA

Substances

RNA
DNA-Directed RNA Polymerases
Phosphoric Diester Hydrolases