用均匀设计优化apoE基因的PCR扩增方案

Optimization of the Amplified Conditions for Human Apo E Gene Fragment by Uniform Design.

由于PCR影响因素多 ,获得理想扩增结果比较困难 ,有必要寻找一种简单有效的方法建立最佳扩增体系 .针对Mg2 +浓度、二甲基亚砜 (DMSO)浓度、变性时间、延伸时间、循环次数等因素进行 4因素 6水平和6因素 10水平均匀设计实验优化apoE基因 2 4 4bp片段的PCR扩增条件 .采用纯化模板和简易模板 ,只需 6～10次实验即可获得特异性、高产率扩增结果 .研究表明 ,将均匀设计用于PCR条件优化以及有关的样品处理、试剂选择等研究方面可以避免盲目性 。

PCR is now one of the key procedure in the field of molecular biology, but too many things affect the PCR result. Sometimes, a lot of PCR optimization experiments must be done before satisfactory results are obtained. So an optimization project with minimum times of test is needed. Uniform Design is what can meet this requirement. For 244 bp fragment in human apo E gene, the optimization of the ampified conditions included concentration of Mg 2+ , concentration of DMSO, denature time, annealing temperature , elongation time, number of cycles based on Uniform Design was reported. The results show that a set of PCR conditions with specific, high yield amplified product can be obtained for purified template or simple treated template by 6～10 times of optimization experiment . The Uniform Design is useful for the optimization of PCR conditions.