微卫星DNA多态性分析在常用近交系小鼠遗传监测中的应用研究

实验动物遗传质量监测的目的是检查该品系的动物是否发生了遗传变异 ,是否混入其它品种、品系动物血缘或发生了错误的交配等 ,以确保检测群体符合该遗传群体的要求。目前国家标准推荐的遗传监测方法主要是生化标记分析法。这一方法的实质是检测同工酶或异构蛋白的变化来推测相应的基因变化 ,因而不可避免地存在着精确度不高、检测位点及反映遗传概貌有限等局限性 ,同时还存在着结果不易分析判读等不足。而微卫星技术具有丰富的可供个体识别标志的微卫星位点 (截止 1999年已知 730 0多个小鼠微卫星位点 ) ,可呈现丰富的可供分析的图带 ,产生…

Microsatellite DNA Polymorphisms in Inbred Strain Mice and Selection as Genetic Monitoring Markers

The aim of genetic monitoring is to checking the genetic contamination within inbred starains, which insures that the strains according with the require of colony . At present the methods based on allozyme biochemistry are the National Standard instructed. methods that using microsatellite DNA would be more useful for genetic monitoring than methods based on allozyme biochemistry because the genome itself is being tested rather than a protein product and a larger portion of the genome can be sampled, and easy to distinguish. methods that using microsatellite DNA had abundant microsatellite loci(over 7300, before 1999) can be identified. Applying enough microsatellite loci will present abundant straps and well polymorphism, which can reflection inherit and variation of roundly genene. In addition, this novel approach allows the rapid, sensitive, convenientand accuracy, even individual identificaton. So we should select microsatellite DNA which is polymorphisms as genetic monitoring markers to determining the strains' origin and genetic background of inbred mice.Untill now Only feasibility has been reported, and in which microsatellite DNA loci have not enough polymorphisms to distinguish genetic differences. Articles on standards and practicality have not been founded in our country. With the optimization of components of reaction buffer and amplificaton parameter, PCR for amplification microsatellite DNA was finally set up. Using the techniques microsatellite DNA can amplified efficaciously. The final concentrations of Mg2+ was 1 . 5-3.0 mmol/L, annealing temperature was 50 ℃-65 ℃. The condition for the PCR amplify were , 94 ℃ for 3min, 30cycles of 94℃ for 30s, 50℃-65℃ for 30s,72℃ for 1min,finally at 72℃ for 1min,then store at 4℃.