种公牛个体识别与鉴定方法研究

本研究采用正交试验法摸索出适于multiple-primer PCR的反应体系中物质成分及其浓度和采用排列组合方法排除干扰引物。结果表明,在10×Buffer(Tris-HCl 100 mmol/L和KCl 500 mmol/L,pH=9.7) 2.5 μL、DDT为8 mmol/L、dNTP为0.8 mmol/L、MgCl₂为2.6 mmol/L、5%甘油、0.25%二甲亚砜和0.5%甲酰胺,Taq酶10 U的25 μL的反应体系中,能够成功为总量为32 pmol/L的11对互不影响的微卫星引物进行multiple-primer PCR。通过这种方法,多个微卫星位点可以同时进行PCR,极大地提高了种公牛个体识别与鉴定的工作效率。

Survey of Method on Individual Identification and Test about Breeding Oxen

This paper studied ingredients and concentration of multiple-primer PCR by orthogonal design and removed interference primer by permutation and combination.Result demonstrated that we successed to set up the system of multiple-primer PCR in reaction system which included 2.5 μL 10×Buffer(100 mmol/L Tris-HCl,500 mmol/L KCl,pH=9.7),8 mmol/L DDT,2.6 mmol/L MgCl2,0.8 mmol/L dNTP,5% glycerol,0.25% dimethy sulfoxide,0.5% formamide and Taq polymerase 10 U.It could provide biology respond for interfering 11 pair primers.So it improved the efficiency of individual identification and superintend the marker of animal husbandry through against fake semen.