墨兰ISSR-PCR反应体系建立及优化  

Establishment of ISSR-PCR Reaction System of Cymbidium Sinense and its Optimization

作　　者：张睿[1] 孙正海[1] 王锦[1] 

机构地区：[1]西南林业大学,云南昆明650224

出　　处：《绿色科技》2014年第3期148-150,共3页Journal of Green Science and Technology

基　　金：国家林业局948项目(铁线莲优良品种与繁育技术编号2008-4-11)

摘　　要：为建立墨兰的ISSR-PCR反应体系,采用正交试验分析了5个因素(Tag酶浓度、Mg2+浓度、模板DNA浓度、引物浓度、dNTP浓度)在4个水平上对ISSR-PCR反应的影响。结果表明:Tag酶浓度为1.6U/25μL、Mg2+浓度为2.4mmol/L、dNTP浓度0.96mmol/L、Primer浓度为0.64μmol/L、DNA浓度3.2ng/25μL是墨兰ISSR-PCR最优反应体系。In order to establish theISSR -PCR reaction systemofISSR -PCR reactionsystem ,this article use-sorthogonalexperiment to analyze the influences of fivefactors including Tagenzyme concentration ,Mg2＋ con-centration ,DNA templateconcentration ,primer concentration ,and dNTP concentrationatfourlevelsof the or-thogonal teston theCymbidiumISSR -PCRreaction system .The results show thatwhen Tag enzyme concen-trationis 1 .6U/25μl ,M g2＋ concentration is 2 .4mmol/L ,dN T Pconcentration is 0 .96 mmol/L ,concentration of primeris 0 .64μmol/L ,DNAconcentrationis 80ng/25μl ,theISSR-PCRreaction system is optimal .

关 键 词：墨兰 ISSR-PCR反应体系 优化 

分 类 号：S682.31[农业科学—观赏园艺]