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Optimization by SRAP-PCR reaction system in rare medicinal herb Panax japonicus C.A.Mey
Author(s): S ONG Jia, ZHANG Shao-Peng, SUN Qiao, LI Tian-Pei, ZENG Wan-Yong, CHEN Ping
Pages: 14-
19
Year: 2015
Issue:
2
Journal: Journal of Wuhan Polytechnic University
Keyword: Panax japonicus; SRAP-PCR; construction; optimization;
Abstract: Panax japonicus C.A.Mey, an important medicinal herb, its wild resources are becoming endangered. Genetic diversity research of P.japonicas is significant for the conservation and sustainable utilization of these re-sources.In this study, optimum SRAP-PCR reaction system for P.japonicus DNA was constructed for the first time.The DNA of rhizome in P.japoncius was abstracted as PCR template.And, the concentration gradient of dNTPs, primer, DNA and Taq polymerase and Tm were assessed respectively to optimize SRAP-PCR reaction sys-tem.An optimal and stable SRAP-PCR system for P.japonicus(Total of 25μl) was as follows:DNA 40 ng, dNTPs 0.2 mmol/L, primer 0.36 umol/L, 10 ×PCR Buffer 2.5 μ,l Taq polymerase 2U , Tm50.Then,24 random prim-ers were chosen for further test.2 couple of primers, Me1/Em5 and Me3/Em1, which can get clear and repeatable bands, were obtained after PCR.Finally, 10 different P.japonicus DNA samples were used to test this method u-sing these 2 primers, which showed that rich polymorphism among P.japoncius plants from different area.Thus, this stable and suitable SRAP -PCR system can be used as an important method of molecular markers analysis for Panax plants genetic diversity studying.
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