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目的建立实验用小型猪微卫星标记的多重PCR体系和进行实验猪群的遗传监测。方法利用3种不同荧光标记的微卫星引物结合ABI3700遗传分析仪测序的方法,通过筛选和优化反应条件,建立可用于实验用小型猪遗传质量控制的稳定的多重PCR反应体系。在此基础上进一步检测实验用小型猪近交群体的遗传变异以验证建立体系的效率。结果筛选出了2组理想的组合:组合1包括SW742、S0228和S0218座位,复性温度58℃和56℃;组合2包括S0155、SW902和S0227三个座位,复性温度为60℃和58℃。组合内不同座位标记不同的荧光染料。还以此检测了实验用小型猪群体中的遗传变异。结论初步建立了中国三种实验用小型猪微卫星标记检测的多重PCR体系,为快速、大通量、准确的小型猪遗传监测提供了初步的技术基础。
Objective To establish a multiplex PCR system for microsatellite markers in experimental miniature pigs and to carry out genetic monitoring of experimental swine herds. Methods Three kinds of microsatellite primers with different fluorescent markers combined with ABI3700 genetic analyzer sequencing were used to screen and optimize the reaction conditions to establish a stable multiplex PCR reaction system for the genetic quality control of miniature pigs in experiment. On the basis of this, we further test the genetic variation of the experimental miniature pig inbred population to verify the efficiency of establishing the system. RESULTS: Two sets of ideal combinations were screened: combination 1 included SW742, S0228 and S0218 loci with refolding temperatures of 58 ° C and 56 ° C; combination 2 comprised three seats of S0155, SW902 and S0227 with refolding temperatures of 60 ° C and 58 ° C . Different combinations within the seat marked different fluorescent dye. The genetic variation in the experimental mini-pig population was also examined. Conclusion The multiplex PCR system for microsatellite marker detection of three experimental miniature pigs in China was initially established, providing a preliminary technical basis for rapid, large-throughput and accurate genetic monitoring of miniature pigs.