Matrix assisted laser desorption/ionization mass spectrometry of enzymatically synthesized RNA up to 150 kDa.
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.
Kirpekar
F
F
Department of Molecular Biology, Odense University, Denmark.
Nordhoff
E
E
Kristiansen
K
K
Roepstorff
P
P
Lezius
A
A
Hahner
S
S
Karas
M
M
Hillenkamp
F
F
D001483:Base Sequence; D012321:DNA-Directed RNA Polymerases / Q000378:metabolism; D007834:Lasers; D013058:Mass Spectrometry; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010727:Phosphoric Diester Hydrolases / Q000378:metabolism; D012313:RNA / Q000032:analysis* / Q000096:biosynthesis / Q000737:chemistry; D017423:Sequence Analysis, RNA
D012313:RNA; D012321:DNA-Directed RNA Polymerases; D010727:Phosphoric Diester Hydrolases