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《Guizhou Agricultural Sciences》 2009-08
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Optimization of ITS-OCR Amplification Conditions of Epimedium acuminatum

LI Mu-dan,SHI Xu,GUAN Ping(College of Life Sciences,Guizhou University,Guiyang,Guizhou 550025,China)  
The DNA extracted from fresh young Epimedium acuminatum leaves by the CATB method was tested in ITS-PCR amplification.The amplification system for ITS analysis is set up by studying the effect of different factors including annealing temperature,Mg2+ concentration,Taq DNA polymerase concentration and dNTP concentration on reaction of ITS-PCR.The optimization conditions for ITS amplification of Epimedium acuminatum are 2.5 μL 10×buffer,1 μL template DNA,1.0 mmol/L MgCl2,1.5 mmol/L dNTP,1.25 U Taq polymerase,0.4 μmol/L ITS 4 and ITS 5.The PCR reaction procedure is 35 cycles under 95℃ 5 min;94℃ 30 s,52℃ 45 s and 72℃ 1 min and then 72℃ 7 min.The optimization conditions for ITS amplification of Epimedium acuminatum can be used in genetic diversity and molecular identification of Epimedium acuminatum.
【Fund】: 贵阳市科学技术计划项目[筑科农合同第(2006)36-2];; 贵州省基金项目[黔科合J字(2008)2267];; 贵州大学研究生创新基金资助项目(校研农2009022)
【CateGory Index】: S567.239
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