Interpretation of oligonucleotide mass spectra for determination of sequence using electrospray ionization and tandem mass spectrometry
Interpretation of oligonucleotide mass spectra for determination of sequence using electrospray ionization and tandem mass spectrometry
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DOI：
10.1021/ac960270t
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发表时间：
1996-07-01
影响因子：
7.4
通讯作者：
McCloskey, JA
中科院分区：
文献类型：
--
作者：
Ni, JS;Pomerantz, SC;McCloskey, JA
描述了用于解释由电喷雾电离产生的多电荷寡核苷酸的碰撞诱导解离的质谱的程序。该方法预期用于在大约15聚体水平及以下的DNA或RNA前对结构完全未知的寡核苷酸进行快速测序。在四极碰撞池中由大量片段化产生的序列相关离子的鉴定主要基于（1）作为质量阶梯传播中的初始步骤的3 ′和5 ′末端残基的识别，（2）已经独立于每个末端构建的重叠核苷酸链的比对，和（3）在排除不正确的候选序列中使用实验测量的分子量。用于序列推导的算法被包含在计算机程序中，该计算机程序需要< 2 s的执行时间。演示了碱基和糖中简单修饰形式的序列定位的解释程序。潜在的混合物的成分直接测序显示使用一个未解决的部分未知的寡核苷酸从核糖体RNA。
Procedures are described for interpretation of mass spectra from collision-induced dissociation of polycharged oligonucleotides produced by electrospray ionization. The method is intended for rapid sequencing of oligonucleotides of completely unknown structure at approximately the 15-mer level and below, front DNA or RNA. Identification of sequence-relevant ions that are produced from extensive fragmentation in the quadrupole -collision cell are based primarily on (1) recognition of 3'- and 5'- terminal residues as initial steps in mass ladder propagation, (2) alignment of overlapping nucleotide chains that have been constructed independently from each terminus, and (3) use of experimentally measured molecular mass in rejection of incorrect sequence candidates. Algorithms for sequence derivation are embodied in a computer program that requires < 2 s for execution. The interpretation procedures are demonstrated for sequence location of simple forms of modification in the base and sugar. The potential for direct sequencing of components of mixtures is shown using an unresolved fraction of unknown oligonucleotides from ribosomal RNA.