A sequencing method for RNA oligonucleotides based on mass spectrometry

被引：33

作者：

Faulstich, K ^{[1
]}

Worner, K ^{[1
]}

Brill, H ^{[1
]}

Engels, JW ^{[1
]}

机构：

[1] UNIV FRANKFURT, INST ORGAN CHEM, D-60439 FRANKFURT, GERMANY

来源：

ANALYTICAL CHEMISTRY | 1997年 / 69卷 / 21期

关键词：

D O I：

10.1021/ac961186g

中图分类号：

O65 [分析化学];

学科分类号：

070302 ; 081704 ;

摘要：

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 C-13-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.

引用

页码：4349 / 4353

页数：5

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