摘要： 利用随机引物扩增多态性(RAPD)分子标记技术研究不同枣树品系之间遗传多态性,建立起一个基于PCR技术的分子遗传标记RAPD分析的优化体系。设置不同的浓度梯度,从dNTPs、随机引物、Taq酶、Mg²⁺、缓冲液Buffer的浓度及模板DNA的质量和用量方面考察,建立了枣树RAPD技术最优体系。结果表明:20μL反应体系组分含量为10×Taq酶Buffer 2μL,Mg²⁺浓度2.0 mmol/L,dNTPs浓度200μmol/L,引物浓度0.2μmol/L,TaqDNA聚合酶浓度0.06 U/μL,DNA模板浓度1.5 ng/μL。最佳扩增程序为94℃预变性4 min,94℃变性30 s,36℃退火40 s,72℃延伸1min,50个循环,最后72℃延伸8 min。

关键词: 枣树, RAPD, 体系优化

Abstract: Genetic diversity between the different Chinese Jujube cultivars was researched by Random Amplified Polymorphic DNA( RAPD) molecular marker technology. The optimized RAPD analysis system of genetic molecular marker on PCR amplification technology must be built. Various concentration of dNTPs, random primer, Taq DNA polymerase, Mg²⁺buffer and DNA template quality and concentrationwas set up. Conclusion of the optimized contents of 20 μ L RAPD reaction system was as follows: 2 μ L 10 ×Taq DNA polymerase Buffer, 2. 0 mmol/L Mg ²⁺, 200 μ mol/L dNTPs, 0. 2 μ mol/L primer,0. 06 U/μ L Taq DNA polymerase and 1. 5 ng/μ L DNA template. The optimized PCR cycle program was as follows: 94℃4 min,94℃30 s,36℃40 s,72℃ 1 min,50 cycles and at final 72℃8 min .

Key words: Chinese Jujube, RAPD, Optimized analysis system

中图分类号: 

• S665.1