部分柿属植物SRAP-PCR反应体系的优化

SRAP技术是一种多态性和信息量丰富的新的分子标记技术,其技术简便、快速,不需预知序列信息,近年来在植物遗传多样性分析、种质鉴定、遗传连锁图的构建以及比较基因组学研究等方面得到广泛应用。为了建立柿属植物SRAP技术体系,对影响SRAP-PCR的Mg2+、dNTPs、Taq聚合酶、引物浓度等因素进行了优化。确定优化的反应体系为:模板DNA30ng,Buffer1×,Mg2+2.5mmol/mL,dNTPs0.2mmol/L,Taq酶1u,引物0.3μmol/L,反应总体积25μL。该体系在柿属植物6种1类型共29个基因型中获得较好的扩增结果,可望在柿属植物起源和进化研究中应用。

Optimization of SRAP -PCR in some Diospyros spp.

SRAP is a new molecular marker which could provide high polymorphism and plentiful information. It is simple and has not the specie-specific character. It had been widely used for genetic diversity, comparing genome analysis and map construction, and so on. The concentrations of Mg2+, dNTPs, Taq DNA olymerase, primers which affect the SRAP-PCR reactions were optimized in order to establish the SRAP molecular marker system in Diospyros spp. The optimum system was as follows: template DNA 30 ng, Buffer 1×, Mg2+ 2.5 mmol/mL, dNTPs 0.2 mmol/L, Taq DNA polymerase 1 u, primer 0.3 μmol/L .The total volume of reaction was 25 μL. Amplications were carriyed out on 29 Diospyros spp. genotypes using this optimum system. The results showed that the system was steady and reliable and would be helpful to study origin and evolution of Diospyros spp.