张美玲, 李巧明. 濒危植物望天树SSR-PCR反应体系的优化[J]. 云南大学学报(自然科学版), 2011, 33(S2): 425-432.
引用本文: 张美玲, 李巧明. 濒危植物望天树SSR-PCR反应体系的优化[J]. 云南大学学报(自然科学版), 2011, 33(S2): 425-432.
shu
ZHANG  Mei ling, LI  Qiao ming. Optimization of SSR-PCR reaction system on an endangerd plant Parashorea chinensis[J]. Journal of Yunnan University: Natural Sciences Edition, 2011, 33(S2): 425-432.
Citation: ZHANG  Mei ling, LI  Qiao ming. Optimization of SSR-PCR reaction system on an endangerd plant Parashorea chinensis[J]. Journal of Yunnan University: Natural Sciences Edition, 2011, 33(S2): 425-432.
shu

濒危植物望天树SSR-PCR反应体系的优化

Optimization of SSR-PCR reaction system on an endangerd plant Parashorea chinensis

    摘要
  • 摘要: 为探索适宜望天树(Parashorea chinensis)的SSR-PCR反应体系，利用L16（45）正交试验设计，对SSR-PCR反应体系中的5个反应组分(dNTPs、引物、DNA模板、Mg2+、Taq DNA聚合酶)进行优化.结果显示，各因素水平的变化对望天树SSR-PCR反应的影响由大到小依次为：模板DNA＞dNTPs ＞引物＞Mg2+＞Taq DNA聚合酶.最终建立了适合望天树SSR-PCR分析的最佳反应体系：在25L反应体系中，10buffer(Mg2+ Free) 2.5L，模板DNA 30 ng， dNTPs 20mmoL/L，引物13moL/L，Mg2+ 50mmoL/L，Taq DNA聚合酶0.5U.利用该优化体系，采用3对不同引物对12份分布于不同地区的望天树材料进行了检验，证明此反应体系具有较好的稳定性和可重复性. 

     

    Abstract: An orthogonal design of L16（45）was applied to optimize the SSR-PCR system on Parashorea chinensis,which employed five major components (dNTPs,primers,template DNA,Mg2+,and Taq DNA polymerase).The results indicated the significance of the effect of five factors on the PCR reaction was in the order of template DNA＞dNTPs ＞primer＞Mg2+＞TaqDNA polymerase.An optimal 25L SSR-PCR reaction system of Parashorea chinensis has been constructed,contained 2.5L 10buffer(Mg2+ Free),30ng template DNA,20mmoL/L dNTPs,13moL/L primers,50mmoL/L Mg2+,0.5U Taq DNA polymerase.To validate the SSR-PCR reaction system above,12 individuals of Parashorea chinensis which distributed in different areas and 3 different SSR primers were employed in this research.The results indicated that the SSR-PCR reaction system is repetitive and steady. 

     

    • HTML全文
    • 参考文献(0)
    • 相关文章
    • 施引文献
  • 资源附件(0)

/

返回文章
返回