摘要
利用SPSS建立正交试验体系,从Taq酶、Mg2+离子、dNTP、模板、引物的浓度5个水平对大麦ISSR反应体系进行优化,确定了适合大麦的快速高效ISSR反应体系。试验结果表明,在25μl反应体系中各反应成分为:0.5 U Taq酶,1μmol/L Mg2+,0.2μmol/LdNTP,0.3μmol/L引物,50 ng模板DNA。这一反应体系的建立对利用ISSR-PCR进行大麦种质资源的分类和新基因资源的挖掘打下了坚实的基础。
Five factors (Taq polymerase, Mg^2+, dNTP, primer, DNA template) of Hordeum vulgare subsp.Vulgare (barley) ISSR amplification system were optimized Using ISSR reaction system to analyze the evolutional relationship of barley.And then the optimum reaction system which contained 0.5 U Taq polymerase, 1 μmol/L Mg^2+, 0.2 μmol/L dNTP, 0.3 μmol/L primer, 50 ng DNA template in a total volume of 25 μl amplification system was established.The optimum reaction system contributed greatly to use the ISSR-PCR technology to classify the quality of Hordeum vulgate subsp.Vulgare and mine the new genes.
出处
《安徽农业科学》
CAS
北大核心
2007年第32期10248-10249,共2页
Journal of Anhui Agricultural Sciences
关键词
大麦
ISSR
SPSS
正交设计
Hordeum vulgate subsp. Vulgare
ISSR
SPSS
Orthogonal design