Smirnov I P, Roskey M T, Juhasz P, Takach E J, Martin S A, Haff L A
PerSeptive Biosystems, 500 Old Connecticut Path, Framingham, Massachusetts, 01701, USA.
Anal Biochem. 1996 Jun 15;238(1):19-25. doi: 10.1006/abio.1996.0243.
A protocol was developed for sequencing oligonucleotides by matrix-assisted laser desorption ionization time-of-flight (MALDI-TOF) mass spectrometry. Oligonucleotides were partially hydrolyzed in separate time-course digestions with 3' --> 5' and 5' --> 3' acting phosphodiesterases in a MALDI-compatible buffer or in the MALDI matrix itself. The partial digests were analyzed by MALDI-TOF mass spectrometry employing an instrument equipped with delayed ion extraction and the sequence was inferred from the mass differences between adjacent peaks. Resolution, mass accuracy, and sensitivity were considerably enhanced with delayed extraction, in comparison with the standard MALDI technique. Much longer lengths of DNA can be unambiguously sequenced with delayed extraction MALDI compared with standard MALDI-MS.
开发了一种通过基质辅助激光解吸电离飞行时间（MALDI-TOF）质谱对寡核苷酸进行测序的方案。寡核苷酸在与MALDI兼容的缓冲液或MALDI基质本身中，分别用3'→5'和5'→3'作用的磷酸二酯酶进行不同时间进程的消化，从而实现部分水解。采用配备延迟离子提取的仪器，通过MALDI-TOF质谱对部分消化产物进行分析，并根据相邻峰之间的质量差异推断序列。与标准MALDI技术相比，延迟提取显著提高了分辨率、质量精度和灵敏度。与标准MALDI-MS相比，使用延迟提取MALDI可以明确测序更长长度的DNA。
