Optimization and Validation of SRAP-PCR Reaction System in Potato

ZHANG Xu,ZHAO Xing-kui,GENG Shu-juan,PENG Suo-tang and DUAN Yong-hong

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Journal of Plant Genetic Resources ›› 2018, Vol. 19 ›› Issue (5) : 993-1000. DOI: 10.13430/j.cnki.jpgr.20180112002
Research Articles

Optimization and Validation of SRAP-PCR Reaction System in Potato

  • ZHANG Xu,ZHAO Xing-kui,GENG Shu-juan,PENG Suo-tang and DUAN Yong-hong
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Abstract

The purpose of this study was to establish the optimal SRAP-PCR reaction system for potato, using the potato genomic DNA as a template and combining the single factor with the orthogonal experimental design, five factors including the concentrations of template DNA, Mg2+, dNTPs, Taq DNA polymerase and primers in SRAP-PCR reaction were optimized, then the optimal SRAP-PCR reaction system for potato was established. The results showed that ,the best SRAP-PCR amplification system is 60 ng template DNA, 1.5 mmol/L Mg2+ , 0.25 mmol/L dNTPs, 0.60 μmol/L each primer and 0.75U Taq DNA polymerasein a 20 μL reaction system for potato. And the order of each factor affecting the result of PCR is: Mg2+ > Taq DNA polymerase > DNA template > primer > dNTPs. The optimized system was verified by 6 potato DNA samples which obtained clear and stable polymorphic bands, it can be used to analyze the genetic diversity and construct genetic map of potato.

Key words

Potato / SRAP-PCR / Optimization of system / Single factor test / Orthogonal desig

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ZHANG Xu,ZHAO Xing-kui,GENG Shu-juan,PENG Suo-tang and DUAN Yong-hong. Optimization and Validation of SRAP-PCR Reaction System in Potato. Journal of Plant Genetic Resources. 2018, 19(5): 993-1000 https://doi.org/10.13430/j.cnki.jpgr.20180112002

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