A Sequencing Method for RNA Oligonucleotides Based on Mass Spectrometry
| Autor: | Konrad Faulstich, Hannelore Brill, Karlheinz Wörner, Joachim W. Engels |
|---|---|
| Rok vydání: | 1997 |
| Předmět: |
Chemical ionization
Chromatography Sequence Analysis RNA Oligonucleotide Chemistry Oligonucleotides Analytical chemistry Nucleic acid sequence RNA Cytidine Mass spectrometry Analytical Chemistry chemistry.chemical_compound Spectrometry Mass Matrix-Assisted Laser Desorption-Ionization Phosphodiester bond Animals Oligoribonucleotides Cattle |
| Zdroj: | Analytical Chemistry. 69:4349-4353 |
| ISSN: | 1520-6882 0003-2700 |
| DOI: | 10.1021/ac961186g |
| Popis: | Synthetic oligoribonucleotides up to 22 bases have been sequenced by observing different kinetics in exonuclease-induced phosphodiester bond hydrolysis and detecting the fragments by matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MS). Common mass spectrometric sequencing methods have disadvantages concerning read length, cost, and specialist instrumentation using RNA as the target molecule because uridine and cytidine have similar masses. Now we are in the position to distinguish U and C by different peak intensities. The method proposed has been verified using specific endonucleases and 13C-labeled nucleotides. This new nongel-based and nonlabeling sequencing strategy offers first RNA sequencing data using the advantages of fast and accurate MALDI-TOF-MS. Preparation steps and measurements are performed in less than 1 h. |
| Databáze: | OpenAIRE |
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