野生猕猴桃DNA的提取及RAPD分析体系的建立

Extraction of DNA and Establishment of RAPD System in Wild Kiwifruit

采用3种不同的方法,即经典CTAB法、改良CTAB法和氯化苄法提取野生猕猴桃基因组DNA,效果均较好,其中改良CTAB法提取的DNA质量最高,DNA降解少,杂质含量少。通过对循环次数、DNA模板用量、DNA聚合酶的种类和浓度等因素的研究,建立了野生猕猴桃RAPD分析体系,从PCR扩增程序和反应体系两个方面对野生猕猴桃RAPD分析体系进行了优化,并验证了优化后的体系,扩增谱带清晰、多态性强,稳定性好。

The three methods for extraction of genomic DNA i. e. classical method, improved CTAB method and benzyl chloride method are practical and efficient. The result showed that the classical method is the best method,because it obtains high-quality genomic DNA with less degradation and im- purity. A number of factors affected the results of RAPD were studied such as cycle numbers,quantity of template DNA and kinds and concentration of Taq DNA polymerase enzyme etc. The optimized RAPD analysis system of the genus kiwifruit was established. Several clear bands with better polymorphism,stability and repeatability could he obtained.