款冬花DNA提取及ISSR体系优化研究  

Genomic DNA Isolation and Optimization for ISSR Reaction System of Tussilago farfara L.

作　　者：贺润丽[1] 畅志坚[2] 樊杰[1] 

机构地区：[1]山西中医学院中药系,太原030024 [2]山西省农业科学院作物遗传研究所,太原030031

出　　处：《中国农学通报》2012年第22期236-241,共6页Chinese Agricultural Science Bulletin

摘　　要：为了从分子水平对款冬种质资源进行鉴定和遗传多样性分析,分别采用SDS法、CTAB法、改良SDS法和改良CTAB法提取款冬叶片基因组DNA,并用改良CTAB法提取的DNA进行ISSR-PCR扩增体系摸索,建立优化的ISSR-PCR反应体系。结果显示:改良SDS法和改良CTAB法都可以提取出高质量的DNA,最优的ISSR-PCR反应条件为:20μL反应体系中含DNA模板约20ng,Taq酶1.0U,Mg2+2.00mmol/L,dNTPs0.20mmol/L,引物0.6μmol/L,1×PCR缓冲液。To investigate the genetic diversity and identification of germplasm resources,the genomic DNA was extracted from the leaves of Tussilago farfara L.by using the SDS,CTAB,modified SDS and modified CTAB methods.The extraction effects of different methods were compared.The subsequent ISSR-PCR condition was explored,using the DNA harvested from modified CTAB methods.The results showed that the modified SDS and CTAB methods could effectively obtain high quality DNA,and the most suitable mixture was as follows：total 20 μL reaction volume containing template DNA 20 ng,1.0 U Taq DNA polymerase,2.00 mmol/L Mg 2＋,0.20 mmol/L dNTPs,0.6 μmol/L pimer,1×PCR buffer.

关 键 词：款冬花 DNA ISSR 反应体系 

分 类 号：R931[医药卫生—生药学]