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：
针对珍稀植物杨叶肖槿ISSR反应的特点，建立了适用于杨叶肖槿遗传多样性研究的ISSR最适反应体系，具体包括：2．0,uL10×Buffer，27.5ng的摸板DNA，2．0uL的dNTP，1U的Pyrobest DNA酶，1．25umol／L的引物；最佳反应程序为94°C预变性5min，然后94°C变性1 min，49°C退火45s，72°C延伸1 min，35个循环；最后72°C延伸10 min，4°C终止反应。应用该优化的反应体系筛选出了10条稳定性强、清晰度高而且表现出一定多态性的ISSR引物，并对杨叶肖槿进行了检测，获得了清晰稳定的扩增图谱。
英文摘要：
To seek a standardizing program of ISSR technique for genetic diversity analysis of Thespesia populnea ,a single factor experiment was designed to optimize ISSR PCR amplification system. The suitable reaction system was obtained,that is 20uL reaction system containing 2.0 uL of 10XBuffer,27.5 ng of to- tal DNA, 2.0 uL of dNTP , 1 U Pyrobest DNA polymerase and 1.25 umol/L of ISSR primer. The profile of ISSR-PCR was an initial denaturation step for 5 min at 94°C ,follow by 35 cycles of 1 min at 94°C ,45 s at annealing temperature 49°C ,1 min at 72°C ,and a final elongation 10 min at 72°C for one cycle,then termi- nation reaction at 4°C . Ten polymorphic primers were screened using this reaction system. Stable and clear amplification patterns were obtained, indicating the ISSR-PCR amplification system was feasible. The fac- tors affecting the amplification of genome DNA of Thespesia populnea were also discussed in the paper. It provides the basis on studies of germplasm resources and genetic diversity of T. populnea.
同期刊论文项目
同项目期刊论文
期刊信息
位置：