樟子松SSR反应体系优化  

Optimization of SSR Reaction System for Pinus sylvestris var. mongolica

作　　者：方攀[1] 张运城[1] 袁虎威[1] 刘玉林[1] 李伟[1] 

机构地区：[1]林木育种国家工程实验室,北京林业大学生物科学与技术学院,北京100083

出　　处：《基因组学与应用生物学》2013年第3期413-419,共7页Genomics and Applied Biology

基　　金：中央高校基本科研业务费专项资金(JD2010-4)资助

摘　　要：从模板DNA、Taq酶含量、dNTP浓度、引物浓度、退火温度、循环次数6个方面,分别设置单因素多水平试验,观察条带的亮度及清晰度,以确定合适的反应体系。然后利用L16(45)正交试验,对DNA模板浓度、Taq酶含量、dNTP浓度、引物浓度这4个因素在4个水平上做进一步优化。建立了樟子松SSR最佳反应体系:25μL的反应体系中包含模板DNA50ng,Taq酶1.0U,dNTP0.3mmol/L,引物0.15μmol/L,10×TaqBuffer(含Mg2+)2.5μL。樟子松SSR反应体系的优化,有助于对樟子松SSR分子标记的研究,为分析樟子松天然群体和人工群体的遗传结构多样性,构建遗传图谱和定位功能基因奠定了基础。In order to research the genetic structure diversity of Pinus sylvestris var. mongolica, SSR reaction sys-tem was optimized in this study. At first, for the sake of determining a suitable reaction system, single factor tests with multi-level were designed respectively under six factors （DNA template, Taq DNA polymerase, dNTP, primer, annealing temperature, cycles）. On the basis of above, the 4 factors which were DNA template, Taq DNA poly-merase, dNTP, Primer were further optimized in 4 levels using L16（4^5） orthogonal test. The obtained optimal reaction system for Pinus sylvestris var. mongolica was as follows：25μL volume of reaction system cotains DNA template 50 ng, Taq DNA polymerase 1.0 U, dNTP 0.3 mmol/L, Primer 0.15μmol/L, 10íTaq Buffer （including Mg^2＋） 2.5μL. The optimization of SSR reaction system for Pinus sylvestris var. mongolica contributes to the development of SSR markers, and lays the foundation for analyzing the genetic structure diversity of Pinus sylvestris var. mongolica in natural populations and artificial populations, constructing the genetic map, and mapping function genes.

关 键 词：樟子松 SSR反应体系 正交试验 

分 类 号：S791.253[农业科学—林木遗传育种]