雷公藤叶片DNA提取及RAPD反应体系建立  

Optimization of RAPD Reaction System on Extraction of Genomic DNA of the leaves of Tripterygium Wilfordii Hook

作　　者：林光美[1] 随粉粉[1] 林楠[2] 郑翔宇[3] 侯长红[4] 

机构地区：[1]福建农林大学作物科学学院,福州350002 [2]福建农林大学生命科学学院,福州350002 [3]福建农林大学食品科学学院,福州350002 [4]福建农林大学经济与管理学院,福州350002

出　　处：《中国农学通报》2010年第1期40-43,共4页Chinese Agricultural Science Bulletin

基　　金：福建省社会发展重点项目(2007Y0047);福建省示范推广重点项目(2006T0014)

摘　　要：筛选并建立雷公藤叶片基因组DNA的Random Amplified Polymorphic DNA技术最优反应体系。采用改良的CTAB法提取叶片总DNA,然后通过对RAPD反应体系中最主要的退火温度、引物浓度、dNTP浓度、DNA模板浓度以及TagDNA聚合酶浓度等5个因子逐个进行单因子优化。结果显示20μlPCR反应体系中,雷公藤叶片基因组DNARAPD最优反应体系为,最佳温度36℃,引物浓度为1.1μmol/L,dNTP浓度1.25mmol/L,DNA模板浓度3790ng/L,TagDNA聚合酶用量为5.625U/L。通过构建该雷公藤叶片基因组的RAPD优化体系,为接下去进一步的多样性分析、遗传图谱构建提供基础。The research aimed to screen out optimum RAPD reaction system on genomic DNA of the leaves of Tripterygium Wilfordii Hook.The improved CTAB DNA extraction method,to extracted the genomic DNA of the leaves of tripterygium wilfordii Hook. The single factor experiment was adopted to select the required 5 annealing temperature,dNTPs concentration,template DNA concentration,Mg2＋ concentration,Taq enzyme concentration and primer concentration. The optimum reaction system for RAPD amplification of the leaves of Tripterygium Wilfordii Hook. Was listed as follows：3790 ng/L DNA,1.1μmol/L primer,1.25mmol/L dNTPs and 5.625 U/L Taq enzyme, the optimum temperature is 56.8℃.All the result could be used in further study about the species diversity of Tripterygium wilfordii Hook. And so forth.

关 键 词：雷公藤 RAPD DNA 反应体系 

分 类 号：S567[农业科学—中草药栽培]