野生濒危掌叶木SRAP-PCR体系的建立与优化  

Establishment and optimization of SRAP-PCR amplification system in Handeliodendron bodinieri

作　　者：李雪萍[1] 苗昌盛[1] 朱恩作[1] 贺春玲[1] 李在留[2] 

机构地区：[1]河南科技大学林学院,河南洛阳471003 [2]广西大学林学院,广西南宁530004

出　　处：《中南林业科技大学学报》2013年第9期18-21,共4页Journal of Central South University of Forestry & Technology

基　　金：国家自然科学基金项目(31060053);河南省自然科学基金项目(62580021);广西自然科学基金项目(2011GXNSFB018016)

摘　　要：以野生濒危植物掌叶木的叶片为材料,通过正交试验与单因素试验相结合的方法,研究了模板DNA用量、Taq DNA聚合酶用量、dNTP浓度、Mg2+浓度、引物浓度等因素对SRAP-PCR反应的影响,建立了适宜于掌叶木SRAP分析的扩增体系,即20μL的反应体系中含模板DNA约20 ng,Mg2+2.0 mmol/L,引物0.6μmol/L,dNTP 0.20 mmol/L,Taq DNA聚合酶1.5 U。该研究为利用SRAP技术分析掌叶木遗传多样性、进行掌叶木种质资源相关研究等奠定了良好的基础。In order to establish an optimal reaction system of SRAP-PCR in Handeliodendron bodinieri, the effects of main elements, such as template DNA concentration, Taq DNA polymerase dosage, dNTP concentration, Mg2＋ concentration, primer concentration on SRAP-PCR reaction were investigated by an orthogonal design and single factor experiments respectively. The optimal reaction system for SRAP analysis in H. bodinieri was obtained. It consisted of 20 ng template DNA, 1 X PCR buffer, 2.0 mmol/L MgC12, 0.20 mmol/L each of dATP, dCTP, dGTP and dTTP, 1.5 unit of Taq DNA polymerase and 0.6 μmol/L primer in a total volume of 20μL. The study lays a good foundation for SRAP analysis in 1t. bodinieri.

关 键 词：掌叶木 SRAP PCR 优化 

分 类 号：S759.95[农业科学—森林经理学] Q948[农业科学—林学]