Mass spectrometric analysis of DNA mixtures: Instrumental effects responsible for decreased sensitivity with increasing mass
Mass spectrometric analysis of DNA mixtures: Instrumental effects responsible for decreased sensitivity with increasing mass
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DOI：
10.1021/ac030127h
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发表时间：
2003-11-01
影响因子：
7.4
通讯作者：
Smith, LM
中科院分区：
文献类型：
--
作者：
Chen, XY;Westphall, MS;Smith, LM
基质辅助激光吸收/电离飞行时间质谱仪在DNA序列分析中显示出取代凝胶电泳法的巨大潜力。然而，这种方法的一个电流限制是，随着DNA分子质量的增加，灵敏度降低。在本研究中，使用等摩尔DNA混合物定量研究了仪器对DNA分子质量分析的影响。结果表明，随着质量的增加，探测效率、探测器饱和度和离子束发散是整个信号强度衰减的原因。虽然本研究侧重于DNA混合物的分析，但观察到的仪器效应同样适用于其他大分子混合物(例如蛋白质、聚合物)。
Matrix-assisted laser absorption/ionization time-of-flight mass spectrometry has demonstrated great potential to replace gel electrophoresis for DNA sequence analysis. A current limitation in this method is, however, the decreased sensitivity with increasing mass of DNA molecules. In the present study, instrumental effects on the mass analysis of DNA molecules were investigated quantitatively using an equimolar DNA mixture. It is shown that detection efficiency, detector saturation, and ion beam divergence account for the entirety of the observed falloff in signal intensity with increasing mass. Although the present study focused upon the analysis of DNA mixtures, the instrumental effects observed apply equally to other macromolecular mixtures (e.g., proteins, polymers).