单雌蓖麻ISSR-PCR反应体系的建立和优化  

Establishment and Optimization of ISSR-PCR Reaction System for Pistillate Castor

作　　者：王亚[1] 郭志强[1,2] 王宏伟[2] 王宙[2] 

机构地区：[1]山西大学生物工程学院,山西太原030006 [2]山西省农业科学院经济作物研究所,山西汾阳032200

出　　处：《山西农业科学》2010年第1期15-18,共4页Journal of Shanxi Agricultural Sciences

基　　金：山西省科技厅攻关项目(20090312006)

摘　　要：以单雌蓖麻提取的基因组DNA为材料,采用单因子试验方法,分别研究了Mg2+浓度、dNTP浓度、引物浓度、Taq DNA聚合酶以及模板DNA用量5种因素对ISSR-PCR反应的影响;建立了适合单雌蓖麻IS-SR-PCR扩增的反应体系,即在20μL反应体系中,10×PCR buffer 2.5μL(Mg2+free),模板DNA 20 ng,2.0mmol/L MgCl2,引物浓度1.0μmol/L,dNTP 0.4 mmol/L,Taq DNA聚合酶1.0 U。Based on the genomic DNA extracted from pistillate castor, the intluencs to ISSR-PCR were optimized and the ef- fect of 5 factors such as Mg2＋ concentration, dNTPs concentration, primer concentration, Taq DNA polymerase dosage and DNA templates dosage on ISSR-PCR amplification were tested using single factor experiment. A reaction system suitable for pistillate castor were. established, that is 20 μ L amplification reactions system containing 10 × PCR buffer 2.5 μ L （Mg2＋free）, 20 ng template DNA, 2.0 mmol/L MgCl2, 1.0 μmol/L primer, 0.4 mmol/L dNTPs, 1.0 U Taq DNA polymerase.

关 键 词：单雌蓖麻 ISSR—PCR 优化 

分 类 号：S565.6[农业科学—作物学]