欢迎您！东篱公司
退出
-
申报数据库
-
立项数据库
-
成果数据库
-
项目获奖数据库
中文摘要：
以小菜蛾（Plutella xylostella）基因组DNA为模板,采用L16（45）正交试验设计方法,建立小菜蛾的ISSR最佳反应体系。通过梯度退火试验,确定不同引物的最适退火温度。优化得到的反应体系（20μL）为：Mg2＋浓度1.5mmol/L、Taq DNA聚合酶2.5U、dNTPs浓度0.2mmol/L、引物浓度1.25μmol/L、模板DNA量20ng。反应程序为：94℃预变性5.0min;94℃变性45.0s,40～61℃（不同引物退火温度各异）退火1.0min,72℃延伸1.5min,40个循环;72℃延伸10.0min;10℃保存。利用所建立的ISSR-PCR反应体系,获得了清晰、重复性好的DNA谱带。
英文摘要：
To establish and optimize ISSR-PCR reaction system for Plutella xylostella, based on the genomic DNA of P. xylostella, the factors influencing ISSR system were explored with the L16 （45 ） orthogonal design. The optimal annealing temperature for ISSR-PCR reaction was proposed by gradient PCR. The optimal conditions for ISSR reaction system （20 μL） were determined as follows： 1.5 mmol/L of Mg2＋ , 2.5U of Taq DNA polymerase, 0.2 mmol/L of dNTPs mixture, 1.25 μmol/L of each primer, and 20ng of template DNA. The reaction program was as followed： initial denaturation for 5.0 min at 94 ℃, 35 cycles of denaturation for 45s at 94 ℃, annealing for 1.0 min at 40--61 ℃, extension for 1.5 min at 72 ℃, with a final extension of 10.0 min at 72 ℃. The clear and reproducible DNA bands were obtained by the established ISSR-PCR reaction system.
同期刊论文项目
同项目期刊论文
期刊信息
位置：