| Optimizing SSR-PCR system of Panax ginseng by orthogonal design |
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| 作者单位： | YANG Tian-tian，MU Li-qiang，WANG Jun School of Forestry of Northeast Forestry University，Harbin 150040，P. R. China |
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| 基金项目： | Foundation project： This research was supported by Department of Wild- life Conservation, State Forestry Administration, P. R. China. |
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| 摘 要： | An orthogonal design was used to optimize SSR-PCR amplification system using Panax ginseng genomic DNA as template. Four levels of five factors （DNA template, Taq DNA polymerase, Mg^2＋, primer, and dNTP） and annealing temperature have been tested separately in this system. The results demonstrated the reaction efficiency was affected by these factors. Based on the results, a stable, productive and reproducible PCR system and cycling program for amplifying a ginseng SSR locus were obtained： 20 μL system containing 1.0 U Taq DNA polymerase, 2.0 mmol·L^-1 Mg^2＋, 0.2 mmol·L^-1 dNTPs, 0.3 μmol·L^-1 SSR primer, 60 ng· μla^-1 DNA template, performed with a program of 94℃ for 5 min, 94℃ for 30 s, annealing at 56.3℃ for 30 s, 72℃ for 1 min, 37 cycles, finishing at 72℃ for 7 min, and storing at 4℃.
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| 关 键 词： | 人参 SSR-PCR系统 正交设计 优化 品种 分子标记 |
| 收稿时间： | 2006-10-24 |
| 修稿时间： | 2006-12-05 |
Optimizing SSR-PCR system of <Emphasis Type="Italic">Panax ginseng</Emphasis> by orthogonal design |
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| Authors: | Yang Tian-tian Mu Li-qiang Wang Jun |
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| Institution: | (1) School of Forestry of Northeast Forestry University, Harbin, 150040, P. R. China |
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| Abstract: | An orthogonal design was used to optimize SSR-PCR amplification system using Panax ginseng genomic DNA as template. Four levels of five factors (DNA template, Taq DNA polymerase, Mg2+, primer, and dNTP) and annealing temperature have been tested separately in this system. The results demonstrated the reaction
efficiency was affected by these factors. Based on the results, a stable, productive and reproducible PCR system and cycling
program for amplifying a ginseng SSR locus were obtained: 20 μL system containing 1.0 U Taq DNA polymerase, 2.0 mmol·L−1 Mg2+, 0.2 mmol·L−1 dNTPs, 0.3 μmol·L−1 SSR primer, 60 ng·μL−1 DNA template, performed with a program of 94°C for 5 min, 94°C for 30 s, annealing at 56.3°C for 30 s, 72°C for 1 min, 37
cycles, finishing at 72°C for 7 min, and storing at 4°C.
Foundation project: This research was supported by Department of Wildlife Conservation, State Forestry Administration, P.
R. China.
Electronic supplementary material is available in the online version of this article at
Biography: YANG Tian-tian (1979–), female, postgraduate student of School of Forestry, Northeast Forestry University, Harbin
150040, P. R. China. |
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| Keywords: | Panax ginseng C A Meyer Orthogonal design SSR-PCR |
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