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《Guihaia》 2008-05
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Establishment and orthogonal optimization of ISSR-PCR amplification system in Salvia miltiorrhiza

LI Rong, WANG Zhe-Zhi( College of Life Sciences, Shaanxi Normal University, Xi’an 710062, China )  
In this paper,the orthogonal design was used to optimize the ISSR-PCR amplification system of Salvia miltiorrhiza in four factors (the concentration of dNTP,Primers,Mg2+ and Taq DNA polymerase) at three levels respectively. Then,based on the optimal ISSR-PCR amplification system,the concentration of template DNA and annealing temperature were proposed by gradient PCR. The results showed that: a suitable ISSR-PCR amplification system of S.miltiorrhiza was established. In a total volume of 20 μL ISSR-PCR amplification system,it contains 1×PCR buffer,200 μmol/L dNTP,1.0 μmol/L primer,1.5 mmol/L Mg2+,1 U Taq DNA polymerase and 20~60 ng template DNA. The optimal annealing temperature for primer UBC 835 is 51.7 ℃.
【Fund】: 国家“十五”科技攻关项目(2004BA721A35)~~
【CateGory Index】: Q943.2
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