利用微卫星多重PCR技术鉴定剑尾鱼RR-B系

目的建立多重PCR技术鉴定选育剑尾鱼RR-B系的方法。方法根据可明显鉴定剑尾鱼RR-B系的6对特异性微卫星引物Msa012、Msa014、Msa033、Msb025、Msd003、Msd051,采用不同的组合方式对RR-B系剑尾鱼和红眼红体的非选育剑尾鱼进行多重PCR扩增。结果引物组合1(Msa014、Msb025、Msd003)和组合2(Msa012、Msa033、Msd051)两个三重PCR反应体系能清楚的扩增各个微卫星座位,且各目的片段之间无干扰,易于识别,引物组合1的排除概率为99.98%,引物组合2的排除概率为99.96%,能准确鉴定剑尾鱼RR-B系。结论本检测方法具有较高的稳定性,可快速准确鉴定剑尾鱼RR-B系。

Identification of Xiphophorus helleri RR-B Strain by Multiplex PCR

Objective To develop a multiplex polymerase chain reaction(PCR) method for identification of RR-B strain of Xiphophorus helleri.Methods 6 pairs of special microsatelites primers,consisted of Msa012,Msa014,Msa033,Msb025,Msd003 and Msd051,were selected to perform multiplex PCR to distinguish RR-B swordtail fish strain from wild swordtail fish.Result Stable amplified product with primer series 1(Msa014,Msb025,Msd003) and series 2(Msa012,Msa033,Msd051) could be obtained.Those amplified segments of multiplex PCR could be used to clearly distinguish the strains from each other.The exclusion probability of primer series 1 was 99.98% and the number of primer series 2 was 99.96%.Conclusion The microsatelites multiplex PCR mothed developed in this study can be used to accurately and rapidly identify the RR-B strain of Xiphophorus helleri.