人参基因组DNA的提取及RAPD-PCR反应条件的优化  

Genomic DNA extraction and optimization of RAPD-PCR conditions of Ginseng

作　　者：周桐[1] 金东淳[1] 伊旭[1] 武立丹[1] 

机构地区：[1]延边大学农学院,吉林龙井133400

出　　处：《延边大学农学学报》2009年第3期189-193,共5页Agricultural Science Journal of Yanbian University

基　　金：国家教育部项目(教外司留[2007]1108);延边大学科技发展项目(延大科合字[2006]20号)

摘　　要：探讨了人参基因组DNA提取方法及RAPD-PCR反应条件的优化.结果表明,用改良的CTAB方法提取的DNA均达到了RAPD-PCR分析的要求;优化的RAPD-PCR反应条件为:在25μL RAPD-PCR反应体系中,模板DNA 20 ng,dNTP 200μmol/L,MgCl21.5mmol/L,10×Buffer 2.5μL,引物浓度0.4μmol/L,Taq DNA聚合酶1.0 U,其余以ddH2O定容至25μL.PCR反应程序为:94℃预变性5 min;94℃45 s,40℃退火1 min,72℃1 min,共40个循环;最后在72℃延伸5 min.Investigated the method of genomic DNA extraction and optimal RAPD--PCR condition. The results showed that the DNA were suitable for RAPD--PCR analysis extrac- ted by CTAB method with some variant;the optimal RAPD--PCR contitions is as follow： in total 25 μL volumes of PCR reaction,containing 20 ng of DNA,200 μmol/L of dNTPs, 1.5 mmol/L of Mge＋ ,2.5 μL of 10× buffer,0.4 μmol/L of primer, 1.0 U of Taq DNA polymerase. The PCR program is：after predenaturing at 94 ℃ for 5 min,followed by 40 cy- cles of denatureing at 94 ℃ for 45 s,annealing at 40 ℃ for 1 min,extension at 72 ℃ for lmin,and final extension at 72 ℃ for 5 min.

关 键 词：人参 DNA RAPD—PCR条件 

分 类 号：S567.51[农业科学—中草药栽培] Q943[农业科学—作物学]