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Optimization of the Conditions of SRAP Molecular Marker Used in Analysis of Fagopyrum tataricum
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作者 王耀文 夏楠 +3 位作者 韩瑞霞 李艳琴 王安虎 蔡光泽 《Agricultural Science & Technology》 CAS 2010年第9期32-36,179,共6页
[Objective] The aim was to investigate the optimal conditions of SRAP molecular marker used in the analysis on Fagopyrum tataricum(L.)Gaertn.[Method]SRAP-PCR amplification system on Fagopyrum tataricum was optimized... [Objective] The aim was to investigate the optimal conditions of SRAP molecular marker used in the analysis on Fagopyrum tataricum(L.)Gaertn.[Method]SRAP-PCR amplification system on Fagopyrum tataricum was optimized by interactive orthogonal design L27(313)in 5 elements(Mg2+,dNTP,Taq DNA polymerase,template DNA and primer)at 3 levels.And the non-denaturing and denaturing PAGE detection methods were compared.The comparative test of DYCZ-24F and DYCZ-20C electrophoresis operating systems was carried out.[Result]The effects of four single-factor(Mg2+,dNTP,Taq DNA polymerase and primer)and two interactions(Mg2+×dNTP,Mg2+×Taq DNA polymerase)on tartary buckwheat SRAP-PCR were significant.An optimal reaction system was established containing 1.5 mmol/L Mg2+,0.2 mmol/L dNTP,1.5 u Taq DNA polymerase,40 ng DNA,0.25 μmol/L primer and 2 μl 10×buffer.Seven samples of tartary buckwheat were amplified using this system,and electrophoresis results showed clear bands,high level of polymorphism and good reproducibility.The PCR products were tested by denaturing and non-denaturing PAGE,and the results showed that the non-denaturing PAGE,DYCZ-24F operating system was more suitable for SRAP analysis.[Conclusion]This study established a foundation for the construction of SRAP genetic map of tartary buckwheat. 展开更多
关键词 Fagopyrum tataricum SRAP-PCR interactive orthogonal design Denaturing PAGE Non-Denaturing PAGE
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