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Electrochemical method for monitoring the progress of polymerase chain reactions using Methylene blue as an indicator |
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| Authors: | Kun Wang Yi-Ping Chen Yun Lei Guang-xian Zhong Ai-lin Liu Yan-Jie Zheng Zhou-Liang Sun Xin-hua Lin Yuan-zhong Chen |
| Affiliation: | 1. Department of Pharmaceutical Analysis, Faculty of Pharmacy, Fujian Medical University, Fuzhou, 350004, China 3. The First Affiliated Hospital of Fujian Medical University, Fuzhou, 350004, People’s Republic of China 2. Fujian Key Lab of Hematology, Fujian Institute of Hematology, Affiliated Union Hospital of Fujian Medical University, Fuzhou, 350000, China 4. The First Affiliated Hospital of Xiamen University, Xiamen, 361003, China |
| Abstract: | We report on the proof-of-principle of an amperometric method to monitor the progress of a polymerase chain reaction (PCR) in real-time. It is based on the finding that the intercalating redox probe Methylene Blue (MB) becomes less easily electrochemically detectable once intercalated into double-stranded DNA (ds-DNA) compared to its free form. This was studied by cyclic voltammetry before and after addition of salmon sperm ds-DNA. Under optimized conditions, the products of the PCR of mitochondrial DNA fragments were quantitatively detected at different stages of amplification cycles. This strategy is potentially cheaper and easier to integrate into a hand-held miniaturized device than fluorescence-based real-time PCR. We report on the proof-of-principle of an amperometric method based on methylene blue which becomes less electrochemically detectable once intercalated into double-stranded DNA compared to its free form to monitor the progress of PCR in real-time. This strategy is potentially cheap and easy to integrate into a hand-held miniaturized device. |
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