酵母菌SRAP-PCR反应体系的优化及遗传多样性分析

从8个正向引物和11个反向引物随机组合成的88个引物对中筛选出14个引物组合,对19株酵母菌进行PCR扩增。采用正交试验设计方法,对SRAP的影响因素进行研究,从Mg2+、dNTPs、DNA浓度、引物、TaqDNA聚合酶设计5因素4水平进行优化试验,以确定PCR电泳分析的最佳条件。14个引物组合共扩增出279条带,其中92.5%呈多态性,相似系数范围为0.60~1.00。

Optimization of SRAP-PCR Reaction System and Genetic Diversity Analysis on Yeasts

Fourteen combinations of SRAP primers screened from 88 SRAP primers combined with 8 orthdromic and 11 reverse primers were used for SRAP amplification of 19 tested yeasts.Orthogonal design was adopted to optimize SRAP amplification system in five factors（Mg2＋,dNTPs,DNA concentrations,primer,Taq DNA polymerase） at four levels respectively,in order to optimize SRAP amplification system.A total of 279 bands were amplified with 14 primers,among which 92.5 % were polymorphic,similarity coefficient ranged from 0.60 to 1.00.