基于单分子操纵技术的有序化测序策略研究-核技术及应用专业论文.docxVIP

Ordered Ordered Sequencing Strategy Based oil the Single Molecule Manipulation Technology Wang Guohua(Nuclear Technology Application) Directed by Prof．Hu Jun Abstraet G-enome sequencing has come true with the development of the DNA sequencing technologies and sequencing strategies．Current sequencing strategies almost based the Shotgun Strategy which was invented in 1980’s by American scientist Sanger．Because of its high randomness，the Shotgun method’S redundancy is very large and assembly work is very difficult and laborious So we put forward new strategy which bases single molecule manipulation technology．It is Ordered Sequencing Strategy． Our strategy elnninates the randomicity of Sanger’S chemical method Firstly, single target DNA sequence is stretched and dissected into small segments orderly, then every segment is amplied by sin羽e molecule PCR experiment and sequenced get sequence infomaation．At last，all segments are combined into consistent sequence in order．Ordered Sequencing Strategy greatly simplify the assembly work， the workload ofthe genome sequencing project is reduced obviously． Single molecule dissection and picking-up technology and single molecule PCR technology are all crucial steps in the whole process．So we also have computed and discussed the rate of sin宙e molecule PCR’S product and its amplification efficiency．After fine computation,we release that if the experiment conditions controlled very well，the rate is low enough to the accomplishment of sequencing project．At the same time，we conside the amplification efficiency maybe is higher than general PCR’S，but this conclusion must be proved by next step experiments Key Words：Genome Sequencing Strategy,Ordered Sequencing，Single Molecule PCR Reaetinn jI苦引 jI苦 引 言 自从DNA双螺旋模型被发现之后，人类对这个组成生命体的最小单元的了 解越来越深入。如今大规模基因组测序已经成为人类了解生命体的一项重要手 段，世界范围内许多模式生物基因组工程都己逐渐完成，我们可以像读书一样阅 读并分析基因组序列。人类基因组工程(Human Oenome Project，HGP)是其中 一项最伟大的工程，它甚至被许多科学家誉为“生物学的圣杯”。 然而随着各个基因组测序工程的完成，科学家们逐渐认识到仅仅能够对某类 基因组实现测序还远远不够，如何才能方便快捷地