叶用莴苣荧光定量PCR体系的优化  

Optimization of Real-time PCR System of Lactuca sativa L.

作　　者：李婷[1] 韩莹琰[1] 范双喜[1] 任月[1] 

机构地区：[1]北京农学院植物科学技术学院,北京102206

出　　处：《中国农学通报》2013年第13期106-113,共8页Chinese Agricultural Science Bulletin

基　　金：北京市自然科学基金项目"热胁迫下叶用莴苣HSP家族基因协同表达机制的研究"(6112005);北京市科技新星计划项目"叶用莴苣耐热机理的研究及耐热品种的选育"(2010B020);现代农业产业技术体系北京市叶类蔬菜创新团队建设专项资金资助-blvt-02

摘　　要：为建立一套最适合叶用莴苣(Lactuca sativa L.)基因表达量研究的荧光定量PCR技术体系,以叶用莴苣S24为试材,采用SYBR Green Ⅰ法,利用叶用莴苣内参基因的2对引物组合18SR1和18SR2,分别在10μL和20μL反应体系条件下进行扩增反应。结果表明,分别在10μL和20μL反应体系条件下内参基因引物组合18SR1的扩增曲线、标准曲线以及溶解曲线等指标都优于内参基因引物组合18SR2;2对内参基因引物组合18SR1和18SR2在10μL反应体系中的扩增曲线、标准曲线以及溶解曲线等指标也均优于20μL反应体系。因此叶用莴苣荧光定量PCR内参基因利用18SR1引物组合,反应体系采用10μL比较合理。In order to establish a method real-time PCR for studying gene expression quantity of Lactuca sative L.,with leaves of Lactuca sativa L.（S24） as material,SYBR Green Ⅰ method had been used,and two pairs of Lactuca sativa L.actin gene primers 18SR1 and 18SR2 were used in 10 μL and 20 μL reaction system conditions amplification reaction.The results showed that primers 18SR1 amplification curve,standard curve and dissolution curve related index were better than primers 18SR2 in 10 μL and 20 μL reaction system conditions.18SR1 and 18SR2 primers combinations in 10 μL reaction system in amplification curve,standard curve and dissolution curve related index were better than 20 μL reaction system.So Lactuca sativa L.18SR1 primers combination and 10 μL reaction system using was more reasonable.

关 键 词：荧光定量PCR SYBR Green Ⅰ 标准曲线 扩增效率 溶解曲线 

分 类 号：S-3[农业科学]