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目的 简化DNA测序方法 ,优化条件 ,以适用于线粒体DNA的测序。方法 应用MaxamGilbert化学断裂反应和连接介导PCR(LigationMediatedPCR ,LMPCR)来测定线粒体DNA的序列。结果 读取的 14 0bp的核苷酸序列 ,输入基因库 ,确定为线粒体DNA重链 80 51～ 8190片段的序列 ,符合率为 70 7% ( 99/14 0 )。结论 该方法改进的成功 ,对进一步进行线粒体DNA氧化损伤图谱分析等研究 ,具有重要意义
Objective To simplify the DNA sequencing method and optimize the conditions for the sequencing of mitochondrial DNA. Methods The sequence of mitochondrial DNA was determined by MaxamGilbert chemical cleavage reaction and Ligation mediated PCR (LMPCR). Results The nucleotide sequence of 14 bp was read into the gene bank and identified as the sequence of 8051 ~ 8190 of the heavy chain of mitochondrial DNA. The coincidence rate was 70.7% (99/14 0). Conclusion The successful improvement of this method is of great significance for further studies on the pattern analysis of mitochondrial DNA oxidative damage