Taranenko N I, Chung C N, Zhu Y F, Allman S L, Golovlev V V, Isola N R, Martin S A, Haff L A, Chen C H
Health Sciences Research Division, Oak Ridge National Laboratory, Tennessee 37831-6378, USA.
Rapid Commun Mass Spectrom. 1997;11(4):386-92. doi: 10.1002/(SICI)1097-0231(19970228)11:4<386::AID-RCM867>3.0.CO;2-N.
The DNA sequence of a single-stranded and double-stranded template was determined. The templates were sequenced using the chain termination method and cycle sequencing method and detected by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. The sequencing products were analyzed successfully without the laborious and expensive methods for removal of the template. Direct sequencing of the double-stranded template was achieved with minimal post-reaction purifications, which could be extremely important for mutation analysis and clinical diagnosis. A systematic study of the mechanisms and kinetics of sequencing reactions was also performed. The details of this analysis and directions for future improvements of the quality of sequencing are presented.
测定了单链和双链模板的DNA序列。使用链终止法和循环测序法对模板进行测序，并通过基质辅助激光解吸/电离飞行时间质谱进行检测。无需采用费力且昂贵的模板去除方法，即可成功分析测序产物。双链模板的直接测序在反应后只需进行最少的纯化，这对于突变分析和临床诊断可能极为重要。还对测序反应的机制和动力学进行了系统研究。本文介绍了该分析的详细情况以及未来提高测序质量的改进方向。
