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首页> 外文期刊>International Journal of Electrochemical Science >tert-Butyl Hydroquinone: an Effective Electrochemical Probe for Sensing Polymerase Chain Reaction and Loop-Mediated Isothermal Amplicons
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tert-Butyl Hydroquinone: an Effective Electrochemical Probe for Sensing Polymerase Chain Reaction and Loop-Mediated Isothermal Amplicons

机译：叔丁基对苯二酚：感应聚合酶链反应和回路介导的等温扩增子的有效电化学探针。

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The quantitative monitoring of nucleic acid amplifications via polymerase chain reaction (PCR) and loop-mediated isothermal amplification (LAMP) are current approaches for high sensitive detection of DNA from pathogenic (micro) organisms. This work is continuous effort of our research group to find inexpensive and readily available DNA-redox intercalating probe for monitoring of amplicons. In this regard, we found tert-butyl hydroquinone (TBHQ), a readily available and low-cost redox molecule popularly used in food additive has good DNA binding ability and it is successful in the quantitative monitoring of DNA amplicons in PCR and LAMP. Our studies revealed that TBHQ has excellent DNA binding abilities and electrochemical behaviors in both PCR and LAMP buffer, and its binding parameters, binding constant, binding site size and diffusion coefficient are suitable for end point detection. The electrochemical end-point detections for monitoring DNA amplifications are established using target genomic DNA as models including λ phage, calf thymus and E. coli and the method is verified and authenticated by traditional methods, such as fluorescence and turbidimetric methods. Besides, we found good recoveries in the determination of target tpc gene spiked in λ phage, calf thymus and E. coli DNA.
机译：通过聚合酶链反应（PCR）和环介导的等温扩增（LAMP）定量监测核酸扩增是目前用于高灵敏度检测病原（微生物）DNA的方法。这项工作是我们研究小组的不懈努力，旨在寻找廉价且易于获得的DNA-氧化还原嵌入探针来监测扩增子。在这方面，我们发现叔丁基对苯二酚（TBHQ）是一种易于获得且低成本的氧化还原分子，广泛用于食品添加剂中，具有良好的DNA结合能力，并且在PCR和LAMP中成功地定量监测DNA扩增子。我们的研究表明，TBHQ在PCR和LAMP缓冲液中均具有出色的DNA结合能力和电化学行为，其结合参数，结合常数，结合位点大小和扩散系数均适合于终点检测。使用靶基因组DNA作为模型（包括λ噬菌体，小牛胸腺和大肠杆菌）建立了用于监测DNA扩增的电化学终点检测，并且该方法已通过传统方法（例如荧光法和比浊法）进行了验证和认证。此外，我们在测定λ噬菌体，小牛胸腺和大肠杆菌DNA中掺入的目标tpc基因时发现了良好的回收率。

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