【摘要】 采用单因素试验设计,以甜高粱基因组DNA为模板,对ISSR反应中的4个主要影响因素:引物浓度、dNTP浓度、Taq DNA聚合酶和Mg2+浓度的用量进行梯度设计优化分析,并对单因素试验结果进行了正交试验设计.研究结果表明,25μL最佳反应体系为:1.0 mmol·L-1MgCl2,1.0 mmol·L-1的dNTP,0.5μmol·L-1的引物,0.5 U的Taq DNA聚合酶.结果显示扩增产物条带清晰明亮、多态性丰富,且特异性强、重复性好,表明本研究所确定的反应体系适用于甜高粱的ISSR分子标记.更多还原

【Abstract】 A single factor experiment with sweet sorghum genomic DNA used as template,was used to determine gradient design optimization analysis for four influencing factors(concentration of primer,dNTP,Taq DNA polymerase and Mg2+)in ISSR reaction. On the basis of the single factor test results,the L9(33)orthogonal test design was then carried out. The optimum content for 25 μL ISSR reaction system was found to be 1.0 mmol·L-1 MgCl2,1.0 mmol·L-1 dNTP,0.5 μmol·L-1 primer,and 0.5 U Taq DNA polymerase. The results showed that the amplified products had clear and bright bands,abundant polymorphism,strong specificity and good repeatability,indicating that the reaction system identified was suitable for ISSR molecular markers of sweet sorghum.更多还原