[Objective] The aim was to optimize the technical procedure of SRAP-PCR in Triticum aestivum L. [Method] The orthogonal design L16（45） was used to optimize SRAP-PCR amplification system of wheat Fengyou 68 on five f...[Objective] The aim was to optimize the technical procedure of SRAP-PCR in Triticum aestivum L. [Method] The orthogonal design L16（45） was used to optimize SRAP-PCR amplification system of wheat Fengyou 68 on five factors （Taq polymerase,Mg2＋,DNA template,dNTPs and primer） at four levels. [Result] Effects of the five factors on SRAP-PCR reaction system were Mg2＋Taq polymerasedNTPsDNA templateprimer. Finally,an optimal SRAP-PCR system was established,that was the total 20 μl reaction system containing 2.0 μl 10×PCR Buffer,2.0 U Taq polymerase,2.0 mmol/L Mg2＋,0.2 mmol/L dNTPs,40 ng DNA template and 0.6 μmol/L primer. [Conclusion] The optimum SRAP-PCR system had provided some technical foundations to conduct SRAP genetic analysis for wheat varieties.展开更多
Camellia oleifera is one of the important oil tree species in south China,and C.oleifera industry is quickly developed with the support of the national policies in recent years.The disorder of C.oleifera varieties is ...Camellia oleifera is one of the important oil tree species in south China,and C.oleifera industry is quickly developed with the support of the national policies in recent years.The disorder of C.oleifera varieties is one of the key issues restricting the development of C.oleifera industry.Because of high polymorphism,good repeatability,less use of DNA and so on,SRAP as a new marker was used in identification of cultivars,analysis of genetic resources and genetic diversity in recent years.In this paper,the orthogonal design was used to optimize SRAP-PCR system for C.oleifera by 5 factors(Mg2+,dNTPs,primer,Taq polymerase,DNA template) and 4 levels,respectively.The data were analyzed by software SPSS V13.0.A suitable SRAP-PCR system(20 μL) was established as: 75 ng DNA template,1.5 mmol·L-1 Mg2+,0.15 mmol·L-1 dNTPs,1U Taq DNA polymerase,0.4 μmol·L-1 primer,1×PCR buffer.The result of optimal SRAP-PCR system will provide a foundation for the identification of C.oleifera cultivars.展开更多
基金Supported by National Key Technology Research and Development Program (2008FY110500-9)~~
文摘[Objective] The aim was to optimize the technical procedure of SRAP-PCR in Triticum aestivum L. [Method] The orthogonal design L16（45） was used to optimize SRAP-PCR amplification system of wheat Fengyou 68 on five factors （Taq polymerase,Mg2＋,DNA template,dNTPs and primer） at four levels. [Result] Effects of the five factors on SRAP-PCR reaction system were Mg2＋Taq polymerasedNTPsDNA templateprimer. Finally,an optimal SRAP-PCR system was established,that was the total 20 μl reaction system containing 2.0 μl 10×PCR Buffer,2.0 U Taq polymerase,2.0 mmol/L Mg2＋,0.2 mmol/L dNTPs,40 ng DNA template and 0.6 μmol/L primer. [Conclusion] The optimum SRAP-PCR system had provided some technical foundations to conduct SRAP genetic analysis for wheat varieties.
文摘Camellia oleifera is one of the important oil tree species in south China,and C.oleifera industry is quickly developed with the support of the national policies in recent years.The disorder of C.oleifera varieties is one of the key issues restricting the development of C.oleifera industry.Because of high polymorphism,good repeatability,less use of DNA and so on,SRAP as a new marker was used in identification of cultivars,analysis of genetic resources and genetic diversity in recent years.In this paper,the orthogonal design was used to optimize SRAP-PCR system for C.oleifera by 5 factors(Mg2+,dNTPs,primer,Taq polymerase,DNA template) and 4 levels,respectively.The data were analyzed by software SPSS V13.0.A suitable SRAP-PCR system(20 μL) was established as: 75 ng DNA template,1.5 mmol·L-1 Mg2+,0.15 mmol·L-1 dNTPs,1U Taq DNA polymerase,0.4 μmol·L-1 primer,1×PCR buffer.The result of optimal SRAP-PCR system will provide a foundation for the identification of C.oleifera cultivars.
