唐古特大黄ISSR-PCR反应条件的优化  

Establishment and optimization of ISSR-PCR reaction conditions for Rheum tanguticum

作　　者：胡延萍[1,2] 谢小龙[1,2] 王莉[1] 杨建[1,2] 李毅[1] 

机构地区：[1]中国科学院西北高原生物研究所高原适应与进化重点实验室,西宁810001 [2]中国科学院研究生院,北京100049

出　　处：《广西植物》2010年第1期112-116,共5页Guihaia

基　　金：国家科技支撑计划项目(2007BAD64B04)~~

摘　　要：利用单因素试验对影响唐古特大黄ISSR-PCR扩增的重要参数进行优化,以期建立其最佳反应条件。结果如下:20μL反应体系包括1.5×PCR buffer(15mmol/LTris-HCl,75mmol/LKCl),1.00mmol/LMgCl2,0.6UTaq DNA聚合酶,0.125mmol/LdNTP,0.5μmol/L引物和30ng模板DNA;引物UBC888适宜的退火温度为57.4℃。ISSR反应条件的建立为利用分子标记技术研究唐古特大黄居群遗传多样性奠定了良好基础。Inter-Simple Sequence Repeats（ISSR）is a good molecular marker for revealing genetic diversity. Reaction system differed in different species,so optimization of ISSR-PCR reaction is very important. Factors which affect the ISSR-PCR amplification,such as the concentration of Mg2＋,Taq DNA polymerase,dNTP,primer and template DNA with different annealing temperatures,were optimized and selected by using the genomic DNA of Rheum tanguticum as material. Optimal PCR（20 μL）mix contained 1.5×PCR buffer（15 mmol/L Tris-HCl,75 mmol/L KCl）,1.00 mmol/L MgCl2,0.6 U Taq DNA polymerase,0.125 mmol/L dNTP,0.5 μmol/L primer and 30ng template DNA. The suitable annealing temperature was 57.4 ℃ for primer UBC888. Establishment of the PCR reaction conditions could favor further studies on the genetic diversity of R.tanguticum by using ISSR molecular marker techniques.

关 键 词：唐古特大黄 ISSR-PCR 优化 

分 类 号：Q943[生物学—植物学]