Britt P F, Hurst G B, Buchanan M V
Chemical and Analytical Sciences Division, Oak Ridge National Laboratory, Tennessee 37831, USA.
J Mass Spectrom. 1996 Jun;31(6):661-8. doi: 10.1002/(SICI)1096-9888(199606)31:6<661::AID-JMS341>3.0.CO;2-Q.
Recent work to apply mass spectrometric methods to DNA analysis has led to the attachment of an electrophore to an oligonucleotide primer, with the purpose of investigating whether the advantages of electron capture ionization (increased ionization efficiency, reduced fragmentation) could be extended to larger molecules, such as Sanger sequence ladders. The stability of the electrophore-modified primers under conditions encountered during matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) was investigated. Four different electrophore labels were successfully attached to the 5' terminus of a 17-base, single-stranded oligodeoxyribonucleotide sequencing primer. The attached electrophore tags are robust under conditions used for sample preparation and MALDI-MS, and little or no fragmentation resulting from loss of the electrophore was observed. While no sensitivity enhancement was observed for the electrophore-labeled DNA, mass spectrometric conditions are discussed under which the electrophore labels could enhance the detection of DNA sequencing ladders.
最近将质谱方法应用于DNA分析的工作已导致将一种电泳体连接到寡核苷酸引物上，目的是研究电子捕获电离的优势（提高电离效率、减少碎片化）是否可以扩展到更大的分子，如桑格测序梯。研究了电泳体修饰引物在基质辅助激光解吸/电离质谱（MALDI-MS）过程中遇到的条件下的稳定性。四种不同的电泳体标签成功连接到一个17个碱基的单链寡脱氧核糖核苷酸测序引物的5'末端。连接的电泳体标签在用于样品制备和MALDI-MS的条件下很稳定，并且未观察到因电泳体丢失而导致的很少或没有碎片化。虽然未观察到电泳体标记的DNA的灵敏度提高，但讨论了质谱条件，在这些条件下电泳体标签可以增强对DNA测序梯的检测。
