酸枣SSR-PCR反应体系优化

Optimization of SSR-PCR Reaction System for Ziziphus acidojujuba

以3个酸枣品系LW1、LW2、LW23为试材,以大枣品种"三星"为对照,采用L16(45)正交实验设计,研究了酸枣SSR-PCR的反应体系,以期为酸枣SSR分子标记研究提供参考依据。结果表明:酸枣SSR-PCR的最佳反应体系为,总体系为20μL,DNA模板量100ng、引物浓度0.5μmol·L^(-1)、Mg2+浓度1.25mmol·L^(-1)、Taq聚合酶量1.5U、dNTPs浓度0.3mmol·L^(-1)。利用该体系,选择引物JSSR250对53个酸枣品系和8个大枣品种的DNA进行扩增,酸枣的扩增产物在140~180bp,具7个等位基因,目标条带清晰,多态性良好。该体系能够应用于酸枣SSR分子标记研究。

Three Zizyphus acidojujuba clones LW1,LW2,LW23 were used as experimental materials,‘Sanxing＇Ziziphus jujuba was as control,by orthogonal experimental design L16（45）,the reaction system of SSR-PCR was established in order to provide the basis in the research method on SSR molecular markers of Zizyphus acidojujuba.The results showed that the optimal systems of SSR（20μL）was DNA templates 100 ng,primer concentration 0.5μmol· L^-1,Mg2＋concentration1.25 mol·L^-1,Taq DNA polymerase 1.5 U,dNTPs concentration 0.3 mmol·L^-1.By the reaction system,using primer JSSR250,DNA amplification of 53 Zizyphus acidojujuba clones and 8 Ziziphus jujuba varieties was carried out,which showed that the target strips were clear and the amplification products ranged from 140-180 bp,with 7 alleles.The optimal reaction systems could be used for the SSR molecular marker development of Zizyphus acidojujuba.