The enhancement of PCR amplification by low molecular-weight sulfones
The enhancement of PCR amplification by low molecular-weight sulfones
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DOI：
10.1016/s0378-1119(01)00621-7
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发表时间：
2001-08-22
期刊：
影响因子：
3.5
通讯作者：
Schutt, CE
中科院分区：
文献类型：
--
作者：
Chakrabarti, R;Schutt, CE
聚合酶链式反应 (PCR) 的 DNA 扩增常常因产量低和特异性低的问题而变得复杂，特别是当目标序列的 GC 含量较高时。优化此类反应的常见方法是向反应混合物中添加少量某些有机化学品，例如二甲基亚砜 (DMSO)、甜菜碱、聚乙二醇和甲酰胺。然而，即使存在此类添加剂，富含 GC 的模板的扩增通常也是无效的。在本文中，我们介绍了一类新型的 PCR 增强化合物，即低分子量砜，它们可有效优化高 GC 模板扩增。我们在此描述了广泛的结构活性研究的结果，其中我们研究了一系列六种不同的砜对 PCR 扩增的影响。我们确定了两种砜，环丁砜和甲基砜，它们是高 GC 模板扩增的特别有效的增强剂，并表明这些化合物通常优于目前使用的两种最有效的 PCR 增强剂 DMSO 和甜菜碱。最后，我们简要讨论了砜官能团在这种增强中可能发挥的作用。 (C) 2001 Elsevier Science B.V. 保留所有权利。
DNA amplification by polymerase chain reaction (PCR) is frequently complicated by the problems of low yield and specificity, especially when the GC content of the target sequence is high. A common approach to the optimization of such reactions is the addition of small quantities of certain organic chemicals, such as dimethylsulfoxide (DMSO), betaine, polyethylene glycol and formamide, to the reaction mixture. Even in the presence of such additives, however, the amplification of GC-rich templates is often ineffective. In this paper, we introduce a novel class of PCR-enhancing compounds, the low molecular-weight sulfones, that are effective in the optimization of high GC template amplification. We describe here the results of an extensive structure-activity investigation in which we studied the effects of a series of six different sulfones on PCR amplification. We identify two sulfones, sulfolane and methyl sulfone, that are especially potent enhancers of high GC template amplification, and show that these compounds often outperform DMSO and betaine, two of the most effective PCR enhancers currently used. We conclude with a brief discussion of the role that the sulfone functional group may play in such enhancement. (C) 2001 Elsevier Science B.V. All rights reserved.