标题:
利用二次正交旋转组合设计优化木瓜SRAP-PCR反应体系
Optimization of Chaenomeles SRAP-PCR Reaction System through the Quadratic Orthogonal Rotatable Combinatorial Design
作者:
臧德奎, 尹长虹, 王延玲, 马燕
关键字:
木瓜属, 二次正交旋转组合设计, PCR体系优化, 数学模型
Chaenomeles; Quadratic Orthogonal Rotatable CombinatorialDesignOptimization of PCR System; Mathematical Model
期刊名称:
《Botanical Research》, Vol.1 No.3, 2012-10-31
摘要:
以木瓜属(Chaenomeles)植物基因组DNA为模板,采用五因素二次正交旋转组合设计(实施),对PCR反应的5个因素(Mg2+、dNTP、引物、Taq DNA聚合酶、模板DNA)进行研究,首次建立了PCR反应的各因素编码值与PCR结果评分值之间的数学模型。利用该模型,建立了PCR反应的最优体系,并进行了验证。最优体系的成功建立表明,该优化方法是稳定可靠的,可以为其他物种的PCR体系优化提供借鉴。
Using Chaenomeles genome DNA as template, the relationship between the five PCR factors (Mg2+, dNTP, primer, Taq DNA polymerase, template DNA) and the PCR results was studied through the quadratic orthogonal rotatable combinatorial design. The mathematical model of the PCR result on the PCR factors was firstly established by software SAS9.0. Based on the model, the optimized amplification system for SRAP-PCR of Chaenomeles was set up and verified. The successful establishment of the optimization system showed that the quadratic orthogonal rotatable combinatorial design are stable and reliable to optimize the SRAP-PCR system, and it could be applied to the other optimization of PCR systems.