太行菊SRAP-PCR反应体系的建立与优化

Establishment and optimization of SRAP reaction system for Opisthopappus Taihangensis(Ling) Shih

相关序列扩增多态性(SRAP)是一种新发展起来的分子标记技术,在太行菊中还未见相关的报道。为建立优化适合太行菊SRAP分析的扩增体系,以太行菊DNA为模板,对影响SRAP-PCR反应体系的5个重要参数设置梯度进行单因素实验分析,以确定最佳的反应条件。经过大量的重复性实验确定了适合太行菊SRPA-PCR反应体系:20μL的反应体系中,模板DNA量50ng,1.25mmol/L的Mg2+浓度,0.5μmol/L的上下游引物,0.2mmol/L的dNTPs以及Taq酶1U。该体系在太行菊个体中均能扩增出清晰稳定的条带,为后续的太行菊遗传多样性分析奠定了基础。

Sequence-related amplified polymorphism （SRAP） is a newly developed molecular marker. Several significant parameters of SRAP-PCR reaction system are studied and optimized by Opisthopappus Taihangensis （Ling） Shih＇s DNA. The stable reaction system is established： 20μL reaction system containing 50ng DNA template, 1.25mmol/LMg2＋, 0.5 μmol/L forward primer, 0.4μmoL/L reverse primer, 0.2mmol/L dNTPs and 1U Taq DNA polymerase. It can be amplificated clear and steady band from this system in the individual of Opisthopappus Taihangensis （Ling） Shih, and laid the foundation for the subsequent research analysis of genetic diversity in Opisthopappus Taihangensis （Ling） Shih.