Ni J, Pomerantz C, Rozenski J, Zhang Y, McCloskey J A
Department of Medical Chemistry, University of Utah, Salt Lake City 84112, USA.
Anal Chem. 1996 Jul 1;68(13):1989-99. doi: 10.1021/ac960270t.
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, from 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 < 2s 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.
本文描述了用于解释通过电喷雾电离产生的多电荷寡核苷酸碰撞诱导解离质谱的程序。该方法旨在对来自DNA或RNA的、结构完全未知的大约15聚体及以下的寡核苷酸进行快速测序。在四极杆碰撞池中通过广泛碎片化产生的与序列相关离子的鉴定主要基于：（1）将3'和5'末端残基识别为质量阶梯传播的初始步骤；（2）对齐从每个末端独立构建的重叠核苷酸链；（3）使用实验测量的分子量来排除不正确的序列候选物。序列推导算法体现在一个执行时间小于2秒的计算机程序中。本文展示了碱基和糖中简单修饰形式的序列定位的解释程序。使用核糖体RNA中未解析的未知寡核苷酸部分，展示了对混合物成分进行直接测序的潜力。
