[Objective] This study aimed optimize sequence-related amplified polymor- phism （SRAP）-PCR system for Paphiopedilum hirsutissimum. [Method] Using P. hir- sutissimum leaf as the material, a single-factor test was ado...[Objective] This study aimed optimize sequence-related amplified polymor- phism （SRAP）-PCR system for Paphiopedilum hirsutissimum. [Method] Using P. hir- sutissimum leaf as the material, a single-factor test was adopted to optimize the fac- tors in SRAP-PCR system, including the concentrations of dNTPs, Mg2＋, Taq poly- merase, DNA template and primers. [Result] The optimized SRAP-PCR system con- tained 2.5 μl of 10xPCR buffer, 0.15 mmol/L dNTPs, 2.0 mmol/L Mg2＋, 0.3 mmol/L primer each, 0.3 U Taq polymerase and 1 μl of DNA template. [Conclusion] This system can amplify clear and repeatable DNA profiles, which can be applied for further study about P. hirsutissimum.展开更多
Part of wild flax species and cultivated species were used as materials for optimizing SRAP-PCR reaction system, and concentration gradient test was made for all factors in order to select and establish the optimal pr...Part of wild flax species and cultivated species were used as materials for optimizing SRAP-PCR reaction system, and concentration gradient test was made for all factors in order to select and establish the optimal program and system fitting for amplifying high diversity, good reproducible and clear bands. This program and system could well meet the requirement of flax SRAP and prove that SRAP marker was feasible for researching genetic polymorphism of flax.展开更多
以丝瓜基因组DNA为模板,采用正交试验设计,对SRAP(sequence related amplified polymorphism,序列相关扩增多态性)反应体系中的4种关键因素(dNTPs、Taq酶、引物、模板)进行优化,建立了适合于丝瓜基因组SRAP分子标记的扩增体系:反应总体...以丝瓜基因组DNA为模板,采用正交试验设计,对SRAP(sequence related amplified polymorphism,序列相关扩增多态性)反应体系中的4种关键因素(dNTPs、Taq酶、引物、模板)进行优化,建立了适合于丝瓜基因组SRAP分子标记的扩增体系:反应总体积10μl,其中含Taq酶1.0U、模板DNA 50.00ng、dNTPs 0.3mmol/L、引物0.30μmol/L。该体系能很好地满足丝瓜基因组SRAP标记的要求,可应用于丝瓜的分子生物学研究。展开更多
基金Supported by Scientific Research Fund for the Introduced Talents in Hechi University(2008QB-N001)~~
文摘[Objective] This study aimed optimize sequence-related amplified polymor- phism （SRAP）-PCR system for Paphiopedilum hirsutissimum. [Method] Using P. hir- sutissimum leaf as the material, a single-factor test was adopted to optimize the fac- tors in SRAP-PCR system, including the concentrations of dNTPs, Mg2＋, Taq poly- merase, DNA template and primers. [Result] The optimized SRAP-PCR system con- tained 2.5 μl of 10xPCR buffer, 0.15 mmol/L dNTPs, 2.0 mmol/L Mg2＋, 0.3 mmol/L primer each, 0.3 U Taq polymerase and 1 μl of DNA template. [Conclusion] This system can amplify clear and repeatable DNA profiles, which can be applied for further study about P. hirsutissimum.
文摘Part of wild flax species and cultivated species were used as materials for optimizing SRAP-PCR reaction system, and concentration gradient test was made for all factors in order to select and establish the optimal program and system fitting for amplifying high diversity, good reproducible and clear bands. This program and system could well meet the requirement of flax SRAP and prove that SRAP marker was feasible for researching genetic polymorphism of flax.
文摘以丝瓜基因组DNA为模板,对SRAP反应中主要五个影响因素d NTP浓度、Taq DNA聚合酶浓度、引物浓度、DNA模板浓度、Mg^(2+)浓度进行优化,建立丝瓜SRAP-PCR反应的体系。根据实验确定的最佳反应体系为25μL SRAP体系。25μL SRAP体系为:75 ng DNA,0.2 mmol/L d NTP,1.25 U Taq酶,0.16μmol/L的单条引物,2.0 mmol/L Mg^(2+),2.5μL 10×Buffer。选用33个丝瓜品种对所确立扩增体系及扩增程序进行验证,检测结果表现为扩增产物条带清晰明亮、亮度高、重复性好,表明本试验所确定的反应体系及反应程序适用于丝瓜的SRAP分子标记。
