利用MPprimer设计引物并优化扩增条件以提高多重PCR效率的实验研究

Improvement of Multiplex PCR Efficiency by Using The MPprimer to Design Primers and Optimizing The Amplification Conditions

多重PCR技术广泛应用于多个研究领域,其中引物设计及扩增条件是提高多重PCR实验效率的关键因素.为探讨优化多重PCR实验的方法,以小鼠5个看家基因为研究对象,使用实验室新近开发的MPprimer程序设计多重PCR引物,并通过改变多种反应条件来优化多重PCR实验.结果表明,MPprimer程序能够设计出理想的多重PCR引物,并且通过对退火温度及延伸时间进行优化,可显著提高多重PCR实验效率,对于提高基因表达的规模化检测能力具有积极的促进作用.

Multiplex PCR was used in many biological fields. Primer design and amplification conditions are critical for enhancing the efficiency of multiplex PCR. In order to improve the multiplex PCR assay, five house-keeping genes of mouse were selected to design primers for multiplex PCR analysis by using the MPprimer program, followed with optimizing the conditions for PCR reactions. Result showed that MPprimer is a valuable tool for multiplex PCR primer design. In addition, by optimizing the conditions for multiplex PCR such as the annealing temperature and extension time, the efficiency of multiplex PCR assay could be significantly improved. This work can be used to advance large-scale gene expression analysis in the post-genome era.