摘要
以大豆为材料，研究了PCR反应体系的主要成分模板DNA浓度、dNTP浓度、引物浓度、Taq酶浓度及退火温度对大豆SSR扩增结果的影响，探索影响SSR扩增结果的各因素的最佳用量及引物的退火温度。结果表明：在试验设计范围内，DNA浓度和dNTP浓度对扩增影响较大，引物浓度在0．05～0．2μmol／L范围内对扩增影响较小，Taq酶浓度对扩增有一定影响，引物要有其扩增适宜的退火温度。确立了适合大豆SSR分子标记研究的优化体系。最终确定总反应体系为20μL，模板DNA20ng，dNTP 200μmol／L，引物0．15μmol／L，Taq酶0．5U，10×Taq Buffer 1．5mmol／L，ddH2O补至20μL。
The SSR amplified results of soybean was studied to optimize several factors applied in SSR technique system, including the concentration of template DNA, dNTP, primer and Taq polymerase. Template DNA and dNTP were found to have distinct effect to amplification in the range of experimentation, primer has a little effect between 0.05 μmol/L and 0.2μmol/L, concentration of Taq polymerase have some effect to amplification, annealing temperature specific to primer pair was also checked. Optimized SSR system for soybean was established： 20 ng of template DNA, 200 μmol/L dNTP,0.15 μmol/L of each primer,0.5 U Taq polymerase and 1.5 mmol/L 10 × Taq Buffer were added to the 20μL PCR mixture.
出处
《华北农学报》
CSCD
北大核心
2007年第5期36-39,共4页
Acta Agriculturae Boreali-Sinica
基金
国家自然科学基金项目(30471094)
山西省科技攻关项目(051017)
山西省高校青年学术带头人项目(200425)
