中文摘要：

定向设计、开发具有特定识别和切割序列的DNA切割试剂（限制性内切酶、金属核酸酶等）在生物技术领域中具有广阔的应用前景．在有关DNA切割的研究工作中，必然涉及切割位点的序列分析，已有的分析方法依赖于Maxam—Gilbert化学断裂法，需要进行同位素标记，容易造成污染；而且操作繁琐，条件控制困难，重复性低，技术要求高；结果依靠放射自显影，灵敏度低而且条带分析有一定的主观性＋我们和RenRui等合作，以Sanger双脱氧法为基本原理，建立了DNA切割位点分析新方法，操作简化，可重复性提高，但是仍然需要进行同位素标记，断裂位点的确定仍有一定的主观性．

英文摘要：

Determining the base sequence of DNA broken site is quite crucial for the study on the cleavage site specificity and mechanism of various natural or synthetic DNA cleavage regents, and on developing novel therapeutic drugs targeting at DNA. The most frequently used method depending on chemical reactions of the Maxam-Gilbert procedure, and the late arising methods used by Rui Ren et al. which were based on Sanger＇s DNA sequencing strategy, all had some deficiencies, either the pollution of radioactive materials, or really complicated and difficult to operate. In the present paper, a new method for DNA cleavage site sequence determination was developed. The fluorescence FAM-labeled primer was annealed to the DNA fragments, which has been cleaved by restriction enzymes or other regents, and extended along the template sequence. The products then loaded onto the polyacrylamide＇ electrophoresis gel of ABI 377 DNA Sequencer. Data was collected and analyzed by using ABI PRISM Data Collection Software and ABI PRISM Sequencing Analysis Software. It is proved to be a credible and simple new approach to determine the base sequence of DNA broken sites .