一种简便快速的制备水稻基因组DNA PCR模板的方法  

A Simple Method for the Rapid Preparation of PCR Template of Oryza sativa Genomic DNA

作　　者：曹燕[1] 王媛[1] 齐进焕[1] 于力[1] 刘秋云[1] 

机构地区：[1]中山大学基因工程教育部重点实验室与生物工程研究中心,广东广州510275

出　　处：《生物技术》2010年第4期30-32,共3页Biotechnology

基　　金：广东省科技计划项目("新型固氮酶基因的转基因植物研究";2008B020100001)资助

摘　　要：目的:发展一种简便快速的制备水稻基因组DNA PCR模板的方法。方法:用枪头捣碎水稻叶片代替液氮研磨法提取水稻基因组DNA作PCR模板,在去污剂SDS和表面活性剂TrionX-100的存在下在沸水中煮沸10min,然后取上清扩增微管蛋白(TubA1)基因内含子。结果:发现在利用煮沸法提取水稻基因组DNA的过程中,用枪头捣碎叶片可代替液氮碾磨,加入0.1%表面活性剂TrionX-100煮沸叶片对制备模板有促进效果,得到了预期的PCR片断。结论:该方法快速、简便、经济,具有良好的重复性与特异性,便于自动化。Objective：To develop a simple method for the rapid preparation of Oryza sativa Indica genomic DNA as PCR template.Method：rice leaf was triturated using pipet tip instead of grounding with liquid nitrogen.Rice leaf was boiled in TE,TE/0.1% SDS and TE/0.1% Triton X-100 respectively and used subsequently for the PCR amplification of an intron of tubulin TubA1 gene .Result：Triturating rice leaves with pipet tips could replace grounding with liquid nitrogen.Subsequent boiling in the presence of 0.1% Triton X-100 for 10 minutes displayed enhancement on PCR amplification.Expected PCR amplicon was obtained with high specificity.Conclusion：This method is simple,rapid,economical,efficient,and apt for automation.

关 键 词：煮沸法 PCR模板 TRION X-100 

分 类 号：Q781[生物学—分子生物学]