【摘要】 目的:寻找一个可用于星星草(Puccinellia tenuiflora(Griseb.)Scribn.et Merr)RAPD-PCR的最适宜的反应体系。方法:利用正交设计法对影响PCR扩增效果的一些因素诸如TaqDNA聚合酶的用量、Mg2+浓度、dNTP以及引物浓度等指标进行筛选和优化。然后,通过引物对该优化结果进行验证。结果:正交设计结果表明,最适宜的PCR反应条件:20μl PCR反应体系中包括10×buffer2μl、模板DNA20ng、dNTP1.6mmol/L、TaqDNA聚合酶0.35U、引物1.0mmol/L、Mg2+1.8mmol/L。验证结果表明,该体系扩增出的条带多且清晰。结论:该研究得出的体系是适合RAPD-PCR的最适宜的反应体系,为今后星星草遗传多样性的研究奠定了基础。更多还原

【Abstract】 Objective:To find a conformable RAPD-PCR reaction system of Puccinellia tenuiflora.Methods:To optimize some factors of RAPD System by the methed of orthongonal design,such as:Taq DNA polymerase dosage,Mg2+,dNTP and primer concentration.Then the random primers were used to validate the result.Results:Orthongonal design result showed that optimistic RCR system of 20μL which contained 10×buffer 2μL,1.6mmol/L dNTP,1.0mmol/L Prime,1.8mmol/L MgCl2,0.35U Taq DNA polymerase and 20 ng template DNA.Validation result showed that the bands of this system expand were more and clear.Conclusion:The system by orthongonal design was the most conformable reaction system for RAPD-PCR,It was established as the good foundation of the study of genetic diversity.更多还原