CHANG Jing,LIU Qingbai,LIU Mingguo.Optimization of SSR-PCR Reaction System for Ziziphus acidojujuba[J].Northern Horticulture,2018,42(12):50-57.[doi:10.11937/bfyy.20174451]
酸枣SSR-PCR反应体系优化
- Title:
- Optimization of SSR-PCR Reaction System for Ziziphus acidojujuba
- 关键词:
- 酸枣; SSR-PCR反应体系; 优化; 正交设计
- 文献标志码:
- A
- 摘要:
- 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20 μL,DNA模板量100 ng 、引物浓度05 μmol·L-1、Mg2+浓度125 mmol·L-1、Taq聚合酶量15 U、dNTPs浓度03 mmol·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140~180 bp,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。
- Abstract:
- 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(20 μL) was DNA templates 100 ng,primer concentration 05 μmol·L-1,Mg2+ concentration 125 mol·L-1,Taq DNA polymerase 15 U,dNTPs concentration 03 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.
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CHANG Jing,LIU Qingbai,LIU Mingguo.Optimization of SSR-PCR Reaction System for Ziziphus acidojujuba[J].Northern Horticulture,2018,42(12):1.[doi:10.11937/bfyy.20174451]
备注/Memo
第一作者简介:常婧(1990-),女,辽宁丹东人,硕士研究生,助理工程师,现主要从事森林培育与经营等研究工作。E-mail:350113050@qq.com.责任作者:刘明国(1964-),男,辽宁朝阳人,博士,教授,现主要从事森林培育与经济林等研究工作。E-mail:liumingguo916@163.com.基金项目:辽宁特聘教授基金资助项目(2012)。收稿日期:2018-02-05