猪不同基因组DNA模板来源和浓度与微卫星位点扩增效果的比较研究

Comparative Study on Different Sources and Concentrations of Swine Genomic DNA Template and the Amplification Effects of Microsatellite Loci

[目的]探讨全部使用猪毛发作为试验材料进行遗传多样性研究的可行性。[方法]以贵州黔北黑猪中的3头成年母猪为试材,分别拔取100、200根毛发和抽取5 ml全血进行DNA提取。以微卫星引物S0225和S0227为引物,设置1.0、2.5、5.0 ng/μl 3种模板浓度进行PCR扩增。[结果]采用2.5、5.0 ng/μl的模板浓度均可获得较好的PCR产物,用于聚丙烯酰胺凝胶电泳对扩增基因有较好的分辨效果。毛发DNA和血样DNA的PCR产物数量和质量均无明显差异。毛发数量与所提取DNA的浓度无直接关系。利用21个全毛样PCR产物进行变性聚丙烯酰胺凝胶电泳,均可观察到清晰的条带。[结论]该研究进一步证明全毛样DNA可作为猪遗传多样性研究的材料。

[Objective] The research aimed to discuss the feasibility of make the genetic diversity research by using swine hair wholly as tested materials.[Method] With 3 adult sows of Qianbei black pigs in Guizhou province as tested materials,100 and 200 hairs were pulled and 5 ml whole blood were taken out to make DNA extraction.With microsatellite primers S0225 and S0227 as primers,3 template concentrations including 1.0,2.5 and 5.0 ng/μl were set up to make PCR amplification.[Result] Using 2.5 and 5.0 ng/μl as template concn.could obtain better PCR products,which had better resolving effects on the amplification genes in polyacrylamide gel electrophoresis.The quantity and quality of PCR products between DNA from hair and DNA from blood samples had no obvious difference.The hair number had no direct correlation with the concentration of DNA.When the PCR products of 21 whole-hair samples were used in the denatured polyacrylamide gel electrophoresis,clear bands could be observed.[Conclusion] The research results further proved that DNA from whole-hair samples could be taken as the materials for swine genetic diversity research.