摘要
在水稻Oryza sativa叶片响应稻纵卷叶螟Cnaphalocrocis medinalis取食后的cDNA-AFLP试验中,对25μL体系中预扩增和选择性扩增的引物浓度、dNTP浓度、镁离子浓度及TaqDNA聚合酶用量4种反应条件影响因子进行不同梯度的优化研究。结果表明,PCR选择性扩增反应时,总反应液25μL体系中引物浓度为0.2μmol/L、dNTP浓度为0.2mmol/L、镁离子浓度为2.0mmol/L、TaqDNA聚合酶用量为13.336n kat/25μL时,可得到更多鲜明可辨的TDFs。
The optimization of eDNA - AFLP selective ampliation reaction system in Oryza sativa L. leaves were studled. Different levels on concentration of primer, dNTP mixture , Mg^2＋ and Taq DNA polymerase were set in this experiment. The results showed that the TDFs bands were clear and more in the following conditions： the primer concentration was 0. 2μmol/L; dNTP concentration was 0. 2mmol/L; Mg^2＋ concentration was 2. 0mmol/L; Taq DNA concentration was 13. 336n kat/25μL.
出处
《山地农业生物学报》
2008年第3期189-194,共6页
Journal of Mountain Agriculture and Biology
基金
国家自然科学基金资助项目(30571128)
贵州省自然科学基金重大资助项目(2002F009)
