槟榔SSR反应体系的建立

Establishment of SSR Reaction System for Areca catechu

以鸡心槟榔为试材,用QIAGEN公司的DNeasy Plant Mini Kit提取基因组DNA,对SSR反应体系进行建立与优化,探讨了槟榔SSR技术中PCR体系的主要成分对扩增结果的影响。最终确定了引物C7549的最佳退火温度为56℃,在PCR反应体系中最佳条件:Mg2+浓度为3.0 mmol/L,dNTPs浓度为0.1 mmol/L,Taq酶量为1.5 U,引物浓度为0.8μmol/L,DNA模板为2.0 ng/μL。利用此反应体系对部分槟榔品种进行PCR扩增并Page胶电泳检测,扩增结果清晰且有较高的多态性,表明该体系适合槟榔的亲缘关系分析。

Genomic DNA was extracted from betelnut （Areca catechu L. ） variety ＂Jixin＂ by DNeasy Plant Mini Kit of QIAGEN Company, the SSR reaction system for bet＇elnnt was established and optimized, and the effects of key components of PCR system in the SSR technology for betelnut on the amplification results were explored. The results showed that the best annealing temperatures for the primer C7549 was 56 ℃, and the optimum 20 μL PCR reaction system included 3.0 mmol/L Mg^2＋ , 0. 1 mmol/L dNTPs, 1.5 U Taq DNA polymerase, 0.8 μmol/L primer, 2.0 ng/μL DNA template. The PCR amplification and Page gel electrophoresis detection on some betelnut cultivars were conducted by using the above reaction system. The amplification results were distinct and had high polymorphism, indicating that this system was suitable for the analysis of betelnut phylogenetic relationship.